There is currently intense interest in developing liquid biopsy applications for cancer screening with a multitude of platforms being implemented to this effect. The contribution and merits of biomarkers types for lung cancer screening will be presented.

This presentation will discuss our key advances towards clinical analyses of extracellular vesicles (EVs): i) development of a high-throughput assay strategy, HiMEX (high-throughput magneto-electrochemical exosome), and ii) assessment of EVs as potent biomarkers for cancer management.

Merkel cell carcinoma (MCC) is a rare and highly metastatic skin malignancy. We compared circulating tumor cell (CTC) detection and characterization in blood samples using the FDA-cleared CellSearch® System and the RosetteSep -DEPArray workflow, to enrich, detect and isolate single CTCs.

In a separate prospective clinical trial, the clinicopathological correlations and prognostic value of PD-L1(+)-CTCs in metastatic breast cancer (MBC) patients were evaluated correlate to survival in MBC.

Although the standard of care for non-small cell lung cancer includes surgery and many options for therapeutics, the response is highly variable and patient specific. Detecting early-stage recurrence would allow clinicians to adapt or change therapy within a curative window, greatly improving outcomes and mortality rates. We are developing a sensitive, simple-to-use, platform based on exogenous labeling that could detect tumor-derived extracellular vesicles in blood and saliva for patient monitoring.

We applied an inertial microfluidic Labyrinth device for highthroughput, biomarker-independent, size-based isolation of CTCs/CTC clusters from patients with metastatic non-small-cell lung cancer (NSCLC). The Labyrinth device recovered heterogeneous CTCs in 100% and CTC clusters in 96% of patients with metastatic NSCLC. The majority of recovered CTCs/clusters were EpCAM, suggesting that these would have been missed using traditional antibody-based capture methods.

ctDNA shows great promise for cancer patient management but there is a need for standardization in its use. A public private partnership was initiated through the Biomarkers Consortium at the FNIH to address this unmet need. This presentation will describe how a pre-competitive alliance came together to better understand the need and acceptable performance characteristics of quality control materials, and provide an update on the intended use and outcomes of the FNIH ctDNA Quality Control Materials project.

As the potential of liquid biopsies for prognostic, predictive or early cancer detection applications grows, so does the demand for technical advances to accompany the burgeoning range of applications. We present new developments that enable targeted re-sequencing for liquid biopsy applications at a fraction of the current cost, while retaining or increasing sensitivity and specificity. Examples for detecting low-level mutations in circulating DNA will be presented.

The quantification of cell-free DNA is important in order to diagnose and interpret a therapeutic response. Digital PCR is known to have increased sensitivity and higher precision over qPCR. The increased sensitivity afforded by dPCR can better identify low levels of circulating cell-free DNA from liquid biopsy samples. Stilla has expanded the multiplexing of the Naica system with 6-color dPCR. 6-color dPCR combined with liquid biopsy samples allows for maximum information output from precious samples.

Longitudinal analysis of circulating tumor DNA has shown promise for monitoring treatment response. However, most current methods lack adequate sensitivity for residual disease detection during or after completion of treatment in patients with nonmetastatic cancer. To address this gap, we have developed targeted digital sequencing (TARDIS) for multiplexed analysis of patient-specific cancer mutations. In this talk, I will share results of TARDIS benchmarking and analysis of plasma samples from early-stage breast cancer patients treated with neoadjuvant therapy.

Cell-free tumor derived DNA (ctDNA) analysis offers the potential for minimally invasive detection of early stage disease as well as minimal residual disease after treatment. However, the utility of ctDNA is currently limited when the tumor burden is low. Here, I will describe novel approaches that leverage knowledge of ctDNA biology and personalized sequencing for sensitive detection even in these challenging settings.

Exosomal proteins and microRNAs are promising biomarkers for cancer liquid biopsy. We have developed an exosome protein microRNA one stop (Exo-PROS) liquid biopsy assay, which enables one-stop capture of tumor-derived exosomes (TEX) and sensitive quantitation of TEX surface proteins and intra-vesicular microRNAs on a single device. The Exo-PROS assay has shown superior sensing performance than ELISA and qRT-PCR. The potential application of Exo-PROS assay has been demonstrated in lung cancer early detection.

The sensitive and accurate quantification of specific cancer biomarkers plays important roles in medical diagnostics and research. We have developed a kinetic fingerprinting approach with dynamically binding probes for the direct, digital, hyper-accurate detection and counting of diverse single biomarker molecules in complex biofluids, including blood serum, urine and in situ, with femtomolar to attomolar limits of detection (LODs). We are currently working to commercialize our technology, termed SiMREPS (single-molecule recognition through equilibrium Poisson sampling).

As cancer care is transitioning to personalized therapies with necessary complementary or companion biomarkers, there is significant interest in determining to what extent non-invasive liquid biopsies reflect the gold standard solid biopsy. We have established an approach for measuring patient-specific circulating and solid cell concordance by introducing tumor touch preparations to the high-definition single-cell analysis workflow for high-resolution cytomorphometric characterization. Patient-specific level of concordance can readily be measured to establish the utility of circulating cells as biomarkers and define biosignatures for liquid biopsy assays.