BME Seminar Series: Rong Fan, Department of Biomedical Engineering, Yale University, Yale Comprehensive Cancer Center

Abstract:

Secreted proteins including cytokines, chemokines, and growth factors represent important functional regulators mediating a range of cellular behavior and cell-cell paracrine/autocrine signaling in tumor microenvironment. Detection of all these proteins is a powerful functional assay of tumor cells as well as other types of cells in a tumor microenvironment. This also represents a new approach to identify deep phenotypes of cancer cells even within the phenotypically pure lineages. However due to the high degree of cellular heterogeneity and, more importantly, polyfunctionality of single cells, it is required to simultaneously measure a large array of functional proteins from each cell and to rapidly assay a large number of single cells in parallel. In this talk, I describe a microchip platform, Proteoplex 1.1, consisting of a large array of sub-nanoliter microchambers integrated with highly miniaturized antibody barcodes for 45-plexed protein detection on thousands of single cells in parallel, which represents a new world record in term of the multiplexing capacity of single-cell protein assay. This platform has been fully validated using human cancer cell lines and compared against the start-of-the-art technologies such as flow cytometric analysis. We further applied this platform to the study of both hematologic cancer and solid tumor. Using a dimensional reduction algorithm, we observed new deep functional phenotypes amongst pure cancer cell lineages in the high-dimensional phenotypic space, which could provide new insights to intra-tumor heterogeneity and evolutionary dynamics. Our technology also has a number of clinician-friendly features such as ease of operation, low sample consumption and standardized data analysis, representing a potentially transformative tool for deep phenotyping of cancer cells in the clinical settings for more accurate diagnosis and monitoring of human cancers.