Circulating Tumor Cells and Liquid Biopsy

2019 Archived Content

The way we think about cancer is about to change. What we used to treat as a random occurrence is becoming both predictable and preventable through an array of emerging circular biomarkers. Through the use of circulating tumor cells, cell-free circulating DNA, exosomes, exomers, oncosomes and other extracellular vesicles, we are now able to understand early stages of cancer and apply molecular profiling to clinical trial selection, patient treatment and minimal residual disease detection. The applications are expanding outside of cancer to include transplant medicine, cardiovascular disease, CNS, autoimmune and infectious disease. Encounter the latest research results and clinical trial data while networking with leaders in the field.

Final Agenda

Scientific Advisory Board

Stefanie Jeffrey, MD, John and Marva Warnock Professor, Surgery; Chief, Surgical Oncology Research, Stanford University School of Medicine

Stuart S. Martin, PhD, Professor, Physiology, Marlene and Stewart Greenebaum NCI Comprehensive Cancer Center, University of Maryland School of Medicine

Catherine Alix-Panabières, PhD, Director, Laboratory of Rare Human Circulating Cells (LCCRH), Pathology and Onco-Biology Department, University Medical Center of Montpellier

Klaus Pantel, MD, Professor and Founding Director, Institute of Tumor Biology, University, Medical Center Hamburg-Eppendorf

Steven A. Soper, PhD, Professor, Micro and Nanofabricated Tools for Biological Discovery and Medical Diagnostics, University of Kansas

Shannon L. Stott, PhD, Assistant Professor, Department of Medicine, Massachusetts General Hospital, Harvard Medical School; BioMEMS Resource Center, The MGH Cancer Center

Monday, March 11

10:30 am Conference Program Registration Open (South Lobby)

11:50 Chairperson’s Opening Remarks

Klaus Pantel, MD, Professor and Founding Director, Institute of Tumor Biology, University, Medical Center Hamburg-Eppendorf

12:00 pm Circulating Tumor Cells in Breast Cancers: Current Clinical Validity and Utility

François-Clément Bidard, MD, PhD, Professor of Medical Oncology, Institut Curie & Versailles St. Quentin University

Numerous clinical data have been collected regarding the clinical validity of CTC count and characterization in both early and advanced breast cancer patients. After having reviewed these data, we will discuss how CTC detection may help customize breast cancer therapy.

12:20 Critical Assessment of the Challenges of Using Blood-Based Biomarkers in Prostate Cancer from a Clinical Point of View

Daniel Danila Daniel C. Danila, MD, Medical Oncology Assistant Attending, Genitourinary Oncology Service, Department of Medicine, Memorial Sloan Kettering Cancer Center

In addition to determining whether circulating tumor cells (CTC) can be used as prognostic biomarkers for patients needing further therapy, current studies have proposed many predictive and pharmacodynamic biomarkers to facilitate the doctor’s ability to optimize and adjust dosage based on their ability to target specific pathways. Significant efforts have been focused on developing and analytically validating predictive biomarkers to select patients most likely to benefit the specific therapy.

12:40 Panel Discussion

1:00 Session Break

1:10 Luncheon Presentation I: Routine Capture and Analysis of Rare Cells for Non-Invasive Diagnostics

Smith_Paul Paul Smith, CEO, ANGLE Biosciences Inc.

This session will focus on key challenges with the capture and analysis of CTCs and CTC clusters for liquid biopsies. Potential applications utilizing epitope independent capture coupled with imaging and highly sensitive, multiplexed molecular techniques will be presented.

1:40 Luncheon Presentation II: Introducing Circulating Stromal Cells and their Clinical Applications

Daniel Adams, Senior Research Scientist, Creatv MicroTech Inc

CellSieve^TMmicrofiltration efficiently collects Circulating Stromal Cells (CStCs) along with CTCs from cancer patient blood. Clinical utility of CStCs includes early detection, companion diagnostics, monitoring treatment response, prognosis and pathogenesis – redefining cancer screening and diagnostics.

2:10 Session Break

2:30 Chairperson’s Remarks

Klaus Pantel, MD, Professor and Founding Director, Institute of Tumor Biology, University, Medical Center Hamburg-Eppendorf

2:40 The Whole Transcriptional Landscape of Circulating Tumors Cells in Metastatic Breast Cancer

Lang_Julie Julie E. Lang, MD, Associate Professor of Clinical Surgery; Director, Breast Cancer Program, Department of Surgery, Keck School of Medicine, University of Southern California

The aim of our study was to evaluate if RNA Seq of circulating tumor cells could serve as a surrogate for biopses of macrometastases. We evaluated treatment opportunities based on circulating tumor cell profiles and tracked them under the selection pressure of systemic therapy in Stage IV breast cancer. RNA Seq of circulating tumor cells may be used to discover molecular alterations that are potentially clinically actionable.

3:10 Large Oncosomes as a Source of Cancer-Specific Circulating Biomarkers

DiVizio_Dolores Dolores Di Vizio, MD, PhD, Division of Cancer Biology and Therapeutics, Departments of Surgery, Biomedical Sciences and Pathology and Laboratory Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center

Extracellular vesicles (EVs) are heterogeneous membrane enclosed structures harboring molecular cargo from the cell of origin. Large oncosomes are a novel subtype of EV that is released by highly migratory cancer cells. Large oncosomes contain abundant tumor-derived cargo, which can be analyzed upon purification from plasma and other biological fluids. Here we show analyses of protein, RNA and DNA in controlled system and patient plasma.

3:40 Molecular Analysis of Circulating Tumor Cells, Extracellular Vesicles and Nucleic Acids as Liquid Biopsy in the Follow-Up of Metastatic Breast Cancer Patients to Stratify Patients for Targeted Therapy

Kasimir-Bauer_Sabine Prof. Dr. Sabine Kasimir-Bauer, Head of Research Laboratory, Department of Gynecology and Obstetrics, University Hospital Essen

The so-called liquid biopsy is discussed to be a surrogate marker for therapy stratification of metastatic breast cancer patients. We compared and analyzed RNA profiles enclosed in circulating tumor cells or extracellular vesicles and performed mutational analysis of cell-free DNA (cfDNA) in plasma samples (Next Generation Sequencing) in the follow-up of the disease to get insights into their feasibility for therapy stratification and to predict therapeutic options.

4:10 An Open, End to End, and Flexible Platform for Scalable CTC Collection, Identification, and Analysis

George_Tad Tad George, PhD, Senior Director of Platform, Applications, RareCyte

RareCyte provides instrumentation and consumables that enable an exquisitely sensitive, accurate, simple, and repeatable workflow from liquid biopsy to single cell isolation. The open and end to end RareCyte CTC workflow will be the focus of this talk.

4:40 Refreshment Break and Transition to Plenary Session

5:00 Plenary Keynote Session (Room Location: 3 & 7)

6:00 Grand Opening Reception in the Exhibit Hall with Poster Viewing

7:30 Close of Day

Day 1 | Day 2 | Day 3 | Download Brochure

Tuesday, March 12

7:30 am Registration Open and Morning Coffee (South Lobby)

8:00 Plenary Keynote Session (Room Location: 3 & 7)

9:15 Refreshment Break in the Exhibit Hall with Poster Viewing

10:15 Chairperson’s Remarks

Martin_Stuart Stuart S. Martin, PhD, Professor, Physiology, Marlene and Stewart Greenebaum NCI Comprehensive Cancer Center, University of Maryland School of Medicine

10:25 Chemotherapy-Induced Pro-Metastatic Changes in Breast Cancer Microenvironment

Oktay_Maja Maja H. Oktay, MD, PhD, Professor of Pathology, The L.G. Koss Division of Cytology, Montefiore Medical Center; Professor of Anatomy and Structural Biology; Director, Clinical Imaging Applications, Integrated Imaging Program, Michael F. Price Center for Genetic and Translational Medicine, Albert Einstein College of Medicine

Chemotherapy may induce pro-metastatic changes within breast cancer microenvironment in both mouse and human breast cancer and subsequent hematogenous cancer cell dissemination to distant sites. Cellular and molecular mechanism involved in chemotherapy-mediated cancer cell dissemination as well as pharmacological inhibitors of this process will be discussed.

10:55 RNA Signatures in CTCs for Precision Cancer Medicine

Miyamoto_David David T. Miyamoto, MD, PhD, Assistant Professor, Radiation Oncology, Harvard Medical School; Department of Radiation Oncology, Center for Cancer Research, Massachusetts General Hospital

CTC analysis is a type of liquid biopsy that enables RNA expression profiling, an analysis not possible with ctDNA. Microfluidic enrichment of CTCs followed by RNA-based digital PCR enables the high-throughput and highly specific detection of CTC molecular signatures in several cancers. These CTC signatures can predict therapeutic responses and may enable the early detection of invasive cancers, thus guiding the precision management of cancer patients.

11:25 Assessing PD-L1 Expression on CTCs and Correlation with Immunotherapy Response

Kulkarni_Rajan Rajan Kulkarni, MD, PhD, Assistant Professor, Dermatology and CEDAR/Knight Cancer Institute, Oregon Health and Science University

PD-1 inhibitors have promising efficacy in several cancers. Circulating tumor cell (CTC) PD-L1 levels could supplement tissue PD-L1 biopsy results by sampling from disseminated tumor sites. Towards establishing CTCs as a screening tool, we developed a protocol to isolate CTCs at high purity and immunostain for PD-L1. Monitoring of PD-L1 expression on CTCs could be an additional biomarker that may help in determining response to immunotherapies.

11:55 Molecular Heterogeneity as Expressed in Cells from Liquid Biopsies using CellSearch and DepArray in Multiple Myeloma

Gross_Steven Steven Gross, PhD, Head, CellSearch Assay Development, Research & Development, Menarini Silicon Biosystems Inc

Exploring the molecular heterogeneity as expressed in cells using CellSearch and DepArray technologies in multiple myeloma and other cancers.

12:25 Session Break

12:35 Luncheon Presentation I : Esophageal Adenocarcinoma: Circulating Tumor Cells in Multimodal Treatment Protocols

Kuvendjiska_Jasmina Jasmina Kuvendjiska, PhD, Surgery, Freiburg University

Our study was designed as a pilot study of the ESOPEC-Trial. We evaluated the presence and morphology of CTCs during the treatment period. The experimental results will be correlated with patients’ overall and relapse-free survival.

1:05 Luncheon Presentation II: Going Further with 3-color Crystal Digital PCR: Characterizing CNV,
Indels and Mutation Linkage

Alexandra Martin, PhD, Application Specialist, Digital PCR, Stilla Technologies

1:35 Refreshment Break in the Exhibit Hall with Poster Viewing

2:05 Chairperson’s Remarks

Soper_Steven Steven A. Soper, PhD, Professor, Micro and Nanofabricated Tools for Biological Discovery and Medical Diagnostics, University of Kansas

2:10 Diagnosing Disease with Rare Circulating EVs: Finding Heterogeneous, Nanoscale Needles in a Nanoscale Haystack

Issadore_David David A. Issadore, PhD, Assistant Professor, Bioengineering & Electrical & Systems Engineering, University of Pennsylvania

Circulating EVs contain a wealth of proteomic and genetic information, presenting an enormous opportunity for liquid biopsy. While microfluidics have been used to successfully isolate cells from complex samples, scaling these approaches for EV isolation has been limited by the low throughput and susceptibility to clogging of nanofluidics. Moreover, the analysis of EV biomarkers is confounded by substantial heterogeneity between patients and within a disease itself. To address these challenges, we developed a multichannel nanofluidic system to analyze crude clinical samples. Using this platform, we isolated EVs, profile the RNA cargo inside of these EVs, and apply a machine learning algorithm to generate predictive panels that could provide useful diagnostics for applications in traumatic brain injury and pancreatic cancer using both murine models and clinical samples.

2:40 Single Cell Morphogenomic and Morphoproteomic Profiling of Liquid Biopsy in Prostate Cancer

Malihi_Paymaneh Paymaneh D. Malihi, PhD Candidate, USC Michelson Center for Convergent Bioscience

Kuhn_Peter Peter Kuhn, PhD, Dean’s Professor of Biological Sciences and Professor of Biological Sciences, Medicine, Biomedical Engineering, and Aerospace and Mechanical Engineering and Associate Director, The Bridge, University of Southern California

Tumor heterogeneity is prevalent in both treatment-naïve localized and end-stage metastasized prostate cancer, and may contribute to the broad range of clinical presentation, treatment response, and disease progression. High Definition-Single Cell Analysis enables morphoproteomic and morphogenomic profiling of single cells from touch preparations of tissue cores as well as liquid samples. HD-SCA platform enables real-time molecular profiling of cells and has the potential to elucidate the origin and evolution of metastatic tumor cells.

3:10 High Precision Isolation and Analysis of Exosomes

Chiu_Daniel Daniel T. Chiu, PhD, A. Bruce Montgomery Professor of Chemistry and Bioengineering, University of Washington

We have recently developed microfluidic and nanofluidic systems for the isolation and analysis of exosomes, offering detailed molecular information with single-exosome resolution. Here, we will describe our technical approach, device performance, and the new information we learned about exosomes as revealed by the new measurements.

3:40 Development and Performance of PlexBio’s IntelliPlex Lung Cancer Panel

Freibe_Peter Peter Friebe, PhD Senior Director Applications and Customer Support – US/EU, PlexBio Co., Ltd.

PlexBio developed the IntelliPlex^TM Lung Cancer Panel as a rapid, comprehensive and cost-effective way to
interrogate patient samples with sensitivities as low as 0.1%. Based on πCode^TM technology, the panel

assesses 36 DNA mutations (KRAS, NRAS, PIK3CA, BRAF and EGFR) and 19 gene variants (ALK, ROS1, RET, NTRK1 and MET).

3:55 Robust, Reliable and Fully Automated Isolation of Free Circulating DNA in Liquid Biopsy Applications

Ockhart_Andrea Andrea Ockhardt, PhD, Product Management, STRATEC Molecular Consumables GmbH

InviMag®DNA Kit enables efficient, standardized, fully automated purification of cfDNA fragments from 4 ml plasma samples. The InviGenius® PLUS simplifies workflows by using a walk-away robotic system for up to 12 samples in parallel. cfDNA is concentrated in 40 µl for analysis.

4:10 St. Patrick’s Day Celebration in the Exhibit Hall with Poster Viewing

5:00 Breakout Discussions in the Exhibit Hall

Parallel Analysis of Circulating Biomarkers in Immunotherapy

Genevieve Boland, MD, PhD, Director, Melanoma Surgery Program, Massachusetts General Hospital; Director, Surgical Oncology Research Laboratories, Massachusetts General Hospital; Assistant Professor, Harvard Medical School; Associate Member, Broad Institute

Clinical application of blood-based biomarkers in melanoma
Unmet clinical needs in blood-based biomarkers
Microvesicle applications in immunotherapy

Clinical Utility and Impact of Liquid Biopsy

Rajan Kulkarni, MD, PhD, Assistant Professor, Dermatology and CEDAR/Knight Cancer Institute, Oregon Health and Science University

Necessary features of technologies/tests for clinical utility
Tumor information that is of clinical relevance
Necessity for standardization

The Importance and Challenge in CTC Culture

Professor Yong-Jie Lu, MBBS, MD, PhD, Professor, Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London

Why is CTC culture important?
What is the challenge?
Does it worth to try it?
Alternative ways to avoid it?
How can we success with CTC culture? The researcher, technology development and the funding supporter/policy marker.

Isolation and Analysis of CTCs

Min Yu, MD, PhD, Assistant Professor, Stem Cell Biology and Regenerative Medicine Member, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California

Downstream analysis of circulating tumor cells
Technologies for CTC isolation
Experimental mouse models for metastasis analysis
Using CTCs as liquid biopsy

6:00 Close of Day

Wednesday, March 13

7:30 am Registration Open and Morning Coffee (South Lobby)

8:00 Plenary Keynote Session (Room Location: 3 & 7)

10:00 Refreshment Break and Poster Competition Winner Announced in the Exhibit Hall

10:50 Chairperson’s Remarks

Jeffrey_Stefanie Stefanie Jeffrey, MD, John and Marva Warnock Professor, Surgery; Chief, Surgical Oncology Research, Stanford University School of Medicine

11:00 Utilizing Extracellular Vesicles for Prediction and Monitoring of Immunotherapy Responses in Melanoma

Boland_Genevieve Genevieve Boland, MD, PhD, Director, Melanoma Surgery Program, Massachusetts General Hospital; Director, Surgical Oncology Research Laboratories, Massachusetts General Hospital; Assistant Professor, Harvard Medical School; Associate Member, Broad Institute

Immune checkpoint inhibitors (ICI) show therapeutic benefit in melanoma. We identified pretreatment and early on-treatment biomarkers that incorporate tumor and host immune status. Our findings suggest that peripheral blood-derived exosomes may serve as a non-invasive biomarker to probe tumor and immune responses to ICI therapy, functioning as both a predictive marker of ICI responsiveness as well as a monitoring tool for tumor persistence and immune activation.

11:30 Molecular Assessment of Circulating Extracellular Vesicles Toward Liquid Biopsy Diagnosis of Gastrointestinal Stromal Tumor and Ewing Sarcoma

Godwin_Andrew Andrew K. Godwin, PhD, Chancellors Distinguished Chair in Biomedical Sciences and Endowed Professor and Director, Molecular Oncology, Professor, Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Deputy Director, University of Kansas Cancer Center

There is growing interest in exploiting circulating extracellular vesicles (EVs), primarily small bioactive nanovesicles (60–120 nm), known as exosomes, as developing sources of biomarkers for diagnosis and therapeutic response. However, EVs are heterogeneous vesicles released by both healthy and tumors cells, with the vast majority of circulating vesicles being derived from normal host cells. To establish their utility as valuable tools for the diagnosis of cancer and therapeutic monitoring of disease progression and response to therapy, we used molecular approaches to profile the proteome of Gastrointestinal Stromal Tumor (GIST)- and Ewing Sarcoma (EWS)-derived EVs, as well as other types of cancer, which provided insights into the oncogenic cargo of these tumor-derived EVs. Using proteins enriched in these different sarcomas, we have developed microfluidic platforms and immunocapture approaches to target tumor-associated EVs and in turn measure exosomal proteins and/or RNAs specific to a cancer subtype to further enhance the specificity of these liquid-based biopsies.

12:00 pm Personalized Detection and Monitoring of Lymphomas Using Circulating Tumor DNA

Ash Alizadeh, PhD, Associate Professor of Medicine, Divisions of Oncology & Hematology, Stanford University School of Medicine

12:30 Enjoy Lunch on Your Own

1:10 Refreshment Break in the Exhibit Hall and Last Chance for Poster Viewing

1:50 Chairperson’s Remarks

Stott_Shannon Shannon L. Stott, PhD, Assistant Professor, Department of Medicine, Massachusetts General Hospital, Harvard Medical School; BioMEMS Resource Center, The MGH Cancer Center

2:00 3D Microfluidic ex vivo Culture of Organotypic Tumor Spheroids to Model Immune Checkpoint Blockade

Russell W. Jenkins, MD, PhD, Member of the Faculty of Medicine, Massachusetts General Hospital Cancer Center, Harvard Medical School

2:30 Patient-Derived Circulating Tumor Cells Inform Mechanisms of Metastasis

Yu_Min Min Yu, MD, PhD, Assistant Professor, Stem Cell Biology and Regenerative Medicine Member, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California

Circulating tumor cells (CTCs) are expected to contain metastasis-initiating cells that can shed light on the mechanisms of cancer metastasis. However, due to limited patient-derived material, the metastatic capability of CTCs has yet to be proved. Using our recently established patient-derived CTC lines, we found that different patient CTC lines demonstrated distinct metastatic tissue tropisms in immunodeficient mice and identified associated pathways to specific organs via RNA-seq and ATAC-seq analysis.

3:00 From Sample-to-Single Cells-to-Answer – An Integrated Microfluidics Approach for Identifying Cancer Cells

Lee_Abraham Abraham “Abe” Lee, PhD, William J. Link Professor and Chair, Department of Biomedical Engineering (BME); Professor, Mechanical & Aerospace Engineering; Director, NSF I/UCRC CADMIM Research Lab: Biomolecular Microsystems and Nano Transducers (BioMiNT), University of California, Irvine

Liquid biopsy is performed by a microfluidic acoustic microstreaming device to isolate and target circulating tumor cells (CTCs) in whole blood. A key bottleneck is to identify the critical subpopulation of cells, often at single cell resolution among billions of cells in circulation. An in vitro perfused vascularized 3D tissue platform can then determine which CTCs can be disseminated to distant tissues through the circulatory system.

3:30 Session Break

3:40 Chairperson’s Remarks

Alix-Panabières_Catherine Catherine Alix-Panabières, PhD, Director, Laboratory of Rare Human Circulating Cells (LCCRH), Pathology and Onco-Biology Department, University Medical Center of Montpellier

3:45 Expansion of Patient-Derived Circulating Tumor Cells from Liquid Biopsy

Lim_Chwee-Teck Chwee Teck (C.T.) Lim, PhD, NUS Society (NUSS) Professor, Department of Biomedical Engineering; Director, Biomedical Institute for Global Health Research and Technology; Founding Principal Investigator, Mechanobiology Institute, National University of Singapore

Personalized therapy in cancer requires monitoring of patients’ individual response to treatment. One approach is to assess drug efficacy on circulating tumor cells (CTCs) obtained from patients’ blood samples (aka liquid biopsy) following ex vivo expansion into CTC clusters. We developed a simple microfluidics-based culture platform that allows co-culture of immune cells with CTCs (obtained after red blood cell lysis of patients’ blood) to promote CTC cluster formation.

4:15 Holy Grail and Big Challenge – Culture of Circulating Tumor Cells

Lu_Yong-Jie Yong-Jie Lu, MBBS, MD, PhD, Professor, Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London

Circulating tumor cells (CTCs) provide a minimally invasive approach to access living cancer cells. The culture of CTCs enables the investigation of their biological features, mechanisms of cancer metastasis and the opportunity for in vitro therapeutic sensitivity tests. Challenge to CTC culture includes their rarity, small proportion of viable cells and unclear favorable growth condition. I will give a brief summary of culture strategies and our own experience.

4:45 Growing Organoids from CTCs: The Importance of CTC Input

Rhim_Andrew Andrew Rhim, PhD, Associate Director for Translational Research Ahmed Center for Pancreatic Cancer Research Assistant Professor of Internal Medicine UT MD Anderson Cancer Center

Liquid biopsies have potential to inform personalized treatments and decision making in patients with cancer. While still in its infancy, the field has exploded, featuring numerous modalities and source material, including circulating tumor cells (CTCs), circulating nucleic acids, exosomes, and others. CTCs, however, hold the greatest promise, as not only do they contain all the tumor-derived material that are in the circulation, but if they could be cultured and propagated ex vivo, the prospect of functional analysis and direct drug combination testing could direct true individualized cancer care. Moreover, since CTCs are in the circulation, serial assay of CTCs, during the throes of treatment, could allow for nimble decision-making to thwart resistance. The major hurdle in the application of CTCs in the clinic is the inability to establish CTC cultures. Our laboratory has hypothesized that the major determinant of culture success is the number of CTCs seeded. This hypothesis is driven in part by evidence suggesting that a sub population of tumor cells retain stem cell like characteristics, called cancer stem like cells. These cells are theorized to be required to initiate tumors and metastases since they possess the capacity for self-renewal. In current protocols, at most 60ml of blood is phlebotomized for CTC isolation. However, we estimate that this yields far too few cancer stem like cells to achieve success in culture. Here we will review the efforts we have undertaken to achieve robust and routine cultures of CTCs, mostly by focusing on dramatically increasing CTC input with less emphasis on epithelial cell purity. In so doing, our preliminary data suggest real promise in our approaches.

5:15 Close of Conference Program

March 11-12, 2025

March 12-13, 2025