The molecular analysis of circulating tumor cells (CTCs) and cell-free tumor DNA (ctDNA) in blood can provide clinically relevant information as “liquid biopsy,” which provides information on tumor biology, early detection of cancer, identification of cancer patients at risk to develop relapse (prognosis), and monitoring tumor evolution, therapeutic targets or mechanisms of resistance. Technical standardization and clinical validation of liquid biopsy assays are essential and currently performed by the Cancer-ID/European Liquid Biopsy Society consortium (www.elbs.eu).

Liquid biopsies could enable cancer treatment response monitoring including the detection of minimal residual disease. I will describe our team's efforts to increase the sensitivity of liquid biopsies to detect low parts-per-million levels of circulating tumor DNA from blood. My talk will include new genomic technologies as well as their application to small clinical studies.

Cell-free DNA (cfDNA) consists of small nucleic acid fragments entering the bloodstream during apoptosis or necrosis. cfDNA fragmentation patterns detected by low-coverage whole genome sequencing can be used to detect the presence of circulating tumor DNA (ctDNA) in a background of cfDNA mostly derived from hematologic cells. This presentation will describe the development of a blood-based, whole-genome, next-generation sequencing (NGS) test to detect early stages of cancer.

Liquid biopsy offers the ability to assess a cancer patient's tumor without a costly and invasive tissue biopsy. The Comprehensive Cancer Profiling with a Next-Generation Liquid Biopsy combines CTC analysis, single-cell genomics, and ctDNA sequencing to empower cellular and genomic characterization of the tumor. This talk will present the capabilities to clinical and translational research and how the next-generation liquid biopsy approach promises to revolutionize the development of future diagnostics and therapeutics.

Real-time identification of tumor-specific molecular alterations is the essence of precision oncology. Technologic advances allow for the detection of mutations, rearrangements, insertions/deletions, copy number alterations, and methylation patterns from peripheral blood DNA with increasing sensitivity. These “liquid biopsies” offer a less invasive, potentially more cost-effective tool to assess prognosis, treatment response, early diagnosis of recurrence, and multicancer early detection. Solutions to promote rapid translation into clinical practice are needed.

Enzymatic tailing of nucleic acids can revolutionize standard molecular biology tools. Dramatic improvements in the areas of isolation, purification, and sample preparation will lead to further advances in multi-omics analyses. 

Introducing a biomimetic approach utilizing cell rolling and adhesion characteristics combined with nanotechnology to enable clinically significant detection and post-capture analysis of circulating tumor cells (CTCs). Join us as we present the integrated BioView-CapioCyte platform featuring a fully automated and standardized CTC enrichment tool with labeling and detection functions for clinical practice

An overall information session on the current development of Liquid Biopsy and EDGC's LB service 'Oncocatch'

The emergence of immunotherapy in oncology requires the discovery, validation and subsequent adoption of robust, sensitive and specific predictive and prognostic biomarkers for daily practice. The use of a liquid biopsy could provide an important complementary or alternative added value to PD-L1 detection in tissue biopsy. In my talk, I will discuss how liquid biopsy could be used in the field of immuno-oncology to predict response or relapse for patients undergoing immune-checkpoint inhibitor therapy.

Liquid biopsies are sources of response biomarkers to measure drug efficacy and monitor patient treatment (including detection of emerging resistance and MRD). Continually developing/improving technologies include CTCs, ctDNA, exosomes, epigenetics, nucleic acid fragments, and micro-RNAs. Analytical validation of the assays is essential and access to large datasets with clinical outcomes and analytics will determine future use. Liquid biopsy-based multi-cancer early detection assays are a recent promising development.

1q21.3 amplification frequently occurs in metastatic melanoma. We determined the utility of 1q21.3 amplification detection of cfDNA to monitor inhibitor checkpoint immunotherapy (ICI) response in metastatic melanoma patients. CfDNA 1q21.3 amplification was analyzed in plasma by multiplex ddPCR from 50 patients receiving ICI. CfDNA from patients who had progressive disease had a higher frequency of 1q21.3 amplification. The specific genes in the 1q21.3 region represent prognostic biomarkers in melanoma patients.

The methylation pattern of plasma circulating-tumor DNA(ctDNA) is the most plentiful indicator of cancer types. The underlying pattern of cancer genomes is global hypomethylation at the intergenic region. This presentation will describe various methods for cfDNA methylation patterns analysis. In particular, we would like to share the results of applying MRE(Methylation-sensitive Restriction Enzyme)-seq, enriching and analyzing the de-methylated regions of cfDNA, and the possibility of using it for multi-cancer early detection.

In recent years, analyses starting from formalin-fixed or fresh frozen tumor tissue samples were complemented by analyses starting from cell-free DNA extracted from patient blood plasma obtained in “liquid biopsies”, requiring validation with a well-defined control standard. In this talk we will cover the difficulties labs face when trying to establish such standards on their own, share our experience with a publicly available standard, present validation results and discuss some caveats.

Extracellular vesicles (EVs) released by tumor cells carry molecular cargo from their cells of origin, making them attractive targets for liquid biopsy development. However, EVs are small and heterogeneous, making them difficult to measure reliably. High-resolution single vesicle flow cytometry (vFC) can directly count, size, and measure the cargo of individual EVs, enabling the identification of EV sub-types that can be targeted as part of liquid biopsy development.

BloodPAC is a public-private consortium developing standards and best practice while operating a data commons to support the liquid biopsy community. In addition to developing standards and aggregating data, BloodPAC works collaboratively with all stakeholders in the field to broaden awareness and implementation of the suggested guidelines and establish a wider chain of feedback and discussion in the community. BloodPAC’s unique approach to collaboration in the field has led to the organization’s success and helps to guide our work into the future. Let's discuss the Consortium's approach, challenges and how BloodPAC enables sharing of information between stakeholders to accelerate progress.

Gene/immune-directed therapies are most effective in cancer subgroups harboring their cognate target. Critical advances in molecular testing involve liquid biopsies, wherein circulating tumor DNA (ctDNA)/circulating tumor cells (CTCs) are analyzed. Liquid biopsies are generally blood based, but can derive from ascites, cerebrospinal fluid, etc. A small tube of blood is exploitable for early cancer diagnosis, identifying molecular abnormalities in shed DNA from multiple metastases, and for monitoring post-treatment changes.

As the potential of liquid biopsies for prognostic, predictive, or early cancer detection applications grows, so does the demand for technical advances that enable the ever-increasing range of applications. We present new technologies allowing highly parallel, PCR-free elimination of wild-type alleles, boosting the ability of all downstream detection methods for detecting point mutations in liquid biopsies and clinical samples. Developments boosting the detection of microsatellite mutations will also be described.

Proteases associated with cancer and other diseases represent an ideal class of biomarkers for liquid biopsy companion (protease inhibitor) diagnostics. Sample to answer assays that require only small volumes (~5-10 ul) of blood, plasma or serum can be carried out in 30-60 minutes using simple electrophoretic formats. Elevated levels of the digestive enzymes trypsin, chymotrypsin, and elastase, and MMP-2, 9 and Cathepsin-S were detected in numerous pancreatic, colorectal and other cancer patient samples.

CNSide™ by Biocept, Inc. is a novel diagnostic assay using enhanced CSF tumor cell recovery, cell labeling, and digital imagery to provide a more accurate, quantitative tumor cell count and molecular characterization of these cells (such as HER2 amplification in breast cancer) to better manage patients with brain metastasis.  Microfluidic cell capture, immunochemistry, FISH, PCR, and NGS of cell-free DNA can be combined.  Dr. Dugan will feature several case studies illustrating how the assay has helped manage patients suffering from leptomeningeal disease (LM).