Abstract

Label-Free Enrichment and Integrated full-length mRNA transcriptome profiling of Single Circulating Tumour Cells from Blood Samples

Jay West, Senior Director of R&D, Fluidigm Corporation

As the precursor to metastatic cancer, circulating tumor cells (CTCs) hold tremendous potential for increasing our understanding of tumor metastasis and advancing personalized medicine. Recent technical advances have enabled the enrichment and genomic characterization of CTCs at the single-cell level and have revealed meaningful and clinically relevant heterogeneity in these rare cells. However, data is lacking for existing methods to link the functional differences of single CTCs to their underlying genomic structure and activity. We have developed an approach that integrates key aspects of biologically based experimentation at the single-cell level using an integrated fluidic circuit (IFC). The Polaris™ system allows for the size-independent selection and isolation of single CTCs from a pre-enriched population based on fluorescent markers. Polaris automatically cultures and exposes the individual cells to a variety of user defined drugs and reagents under environmentally controlled conditions or processes them directly for mRNA seq. If desired, an extracellular matrix (ECM) can be deposited into each of the single-cell culture chambers. Cell activity and phenotype can be monitored in situ in response to treatment conditions, followed by preparation of the mRNA transcriptome for RNA seq analysis. Because the entire workflow following pre-enrichment is integrated on a single microfluidic device, little manipulation of the single cell is required, helping preserve the mRNA signature of each cell.