The Single Cell Analysis Core develops innovative technologies for quantitative single-cell assays and facilitates the research proposed in Projects 1, 2 and 3 of the DFCI PS-OC. More specifically, this core will take advantage of the most recent advances in microfluidic single-cell handling to establish five single-cell analysis platforms.
1. Single-cell secretomic profiling;
2. Single-cell phospho-protein profiling;
3. Single-cell whole genome amplification for genome-wide functional analysis;
4. Single-cell epigenomic analysis;
5. Laser scanning cytometry for high-content single-cell analysis.
In addition, the core will be equipped with the state-of-the-art single cell analysis tools such as flow cytometer and motorized fluorescence microscope. These platforms together will enable a suite of functional evaluation ranging from genomics, epigenomics to proteomics of single primary tumor cells and examine phenotypic variability, functional state of activation and/or the response to drugs in heterogeneous populations of cells. For example, they will provide approaches to measure phospho-profiles of drug-sensitive pathways (e.g. EGFR), downstream signaling cascades (e.g. AKT and MAPK pathways), cell-cycle phases, and apoptotic response at the single cell level. Special emphasis is given to quantification of parameters for the dynamics of tumor cell proliferation and death (e.g. during drug treatment with varying concentration). This core contributes to the PS-OC by providing quantitative measurements that parameterize “cellular fitness"(as a convolution of cell cycle and apoptosis dynamics) and signaling events in colorectal/gliomaneurospheres/leukemia lines of the relevant genotypes; these measurements are then inserted into the modeling framework of the PS-OC. To achieve these core's objectives (i.e. to establish a facility dedicated to providing quantitative measurements of cancer growth, signaling, and response to drugs in single-cell resolution for animal and cell line models of tumorigenesis), we 1) establish validated quantitative measurements of proliferation and death rates of primary tumors and associated molecular signatures; 2) provide single-cell resolution of the variability of cellular signaling response during varying conditions (such as exposure to growth factors, cytokines, drugs, radiation therapy etc); and 3) establish systematic protocols to test the drug response of primary tumor cells.