Abstract 234: Snapshot of MAP kinase and related signal transduction pathways from biopsied cells using a novel non-optical assay technology - a proof of concept study

Abstract

Abstract Aberrant activity of the MAP and PI3 kinases is implicated in many forms of cancer. The ability to gather quantifiable information on the activation-state of these kinases from limited in-vitro and in-vivo tumor samples would accelerate drug development and ultimately, the treatment of cancer. Here non-optical, AMMP (acoustic membrane microparticle) technology is used to quantitate the activity state of multiple kinases including EGFR, c-RAF, MEK, ERK, AKT, p38 and JNK in a single assay plate. Lysates from multiple unstimulated tumor cell lines were compared with those from the same cell lines specifically stimulated with ligands to several well-known surface receptors for expressed changes in their phosphorylation states. The data show that the AMMP assay technology can be used to monitor the MAP Kinase pathway activation from EGFR stimulation through ERK phosphorylation following EGF stimulation. Additionally, the activation states of PI3 kinase, p38 and JNK were determined from the same lysate samples. Subsequently, assays were developed to monitor kinase dimerization (e.g., MEK-ERK) by rearranging the pairing of antibodies from the kinase activity assays described above. Using fewer than 2000 cells per assessment, multiple kinases were measured, in their native states, in a single assay using AMMP technology. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 234. doi:1538-7445.AM2012-234