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Single-Cell Biophysics: Measurement, Modulation, and Modeling
Poster Abstracts
99
2-POS
Board 1
Analyzing the Role of CXCR4 in Metastatic Prostate Cancer through Single-Cell Analysis
Joseph Bae
, Anna Sandstrom Gerdtsson, Peter Kuhn.
University of Southern California, Los Angeles, CA, USA.
Metastasis to the bone marrow and other sites comprises the most lethal phase of prostate cancer
progression. Circulating tumor cells (CTCs) are present in the blood at a low concentration, and
play a role in the mechanism of metastasis. Homing and retention of cancer cells from the
primary tumor to the bone marrow niche is thought to be mediated by the CXCR4 chemokine
receptor and its ligand, CXCL12. The High-Definition Single-Cell Analysis (HD-SCA) assay
utilizes fluorescent staining and high-throughput microscopy to identify and characterize CTCs
among the patient’s leukocyte population in blood and bone marrow samples based on
expression of cytokeratin (epithelial cell marker), DAPI (nuclear stain) and CD45 (white blood
cell marker). Additionally, this novel platform is useful in identifying various subpopulations of
CTCs that are not characterized by other similar assays. The HD-SCA assay can also be
expanded upon to accommodate analysis of additional biomarkers of interest by introducing new
assays to the platform. CTCs that have been identified using the HD-SCA assay can then be
individually isolated and further analyzed through genomic or proteomic profiling. In this study,
we have used prostate cancer PC3 cells to develop a HD-SCA compatible fourth color assay for
identifying the expression of CXCR4. The assay will be used to elucidate the role of CXCR4 in
the homing of tumor cells to metastatic sites such as the bone marrow, and to characterize the
relative levels of expression among tumor cells in primary, circulatory, and metastatic
compartments.