High-Throughput Analysis of MAPK and JAK-STAT Signaling in CT26 Tumors Using a Combination of Immunophenotyping and Phospho-Flow Cytometry

AACR 2018 -- The development of robust platforms that enable phospho-protein-based analysis of cell signaling in the tumor microenvironment is pivotal for small molecule drug development. Currently lacking are reliable applications that can simultaneously measure multiple phospho-proteins in both tumor cells and immune cells ex vivo. This can be crucial because many signaling pathways are commonly involved in both pro- and anti-tumor responses in these two cellular compartments. Labcorp has validated a phospho-flow cytometry-based platform that measures the phospho-protein levels of multiple signaling proteins in separate and distinct subsets simultaneously, which can include solid tumor-derived immune subsets and tumor cells. The Labcorp phospho-flow platform can be applied to the analysis of treated cell lines and heterogeneous tissue-derived cell cultures in vitro as well as the ex vivo analysis of inflamed tissue from pharmacology studies. In this presentation, we demonstrate how these services can be used to analyze MAPK and JAK-STAT pathway-associated kinases in vitro in cultured MV-4-11 acute myeloid leukemia (AML) cells and activated murine splenocytes, as well as ex vivo in both tumor-derived CD8+ T cells and tumor cells using the CT26 model for colorectal carcinoma.