PIK3CA Mutational Analysis to Reduce Sample Amount, Reduce Cost, and Increase Throughput to Broadly Assess Acquired Mutational Status

The phosphoinositide-3 kinase (PI3K)/AKT signaling network is one of the two most commonly mutated pathways identified in human cancers and the most frequently mutated pathway in breast cancer. Somatic alterations including PIK3CA mutations are the most common genetic alteration of this pathway; 80% or more occur within the helical (E542K, E545K) and kinase (H1047R) domains of p110. Such mutations confer increased catalytic activity for the generation of the second messenger phosphatidylinositol (3,4,5) - triphosphate (PIP3) and downstream pathway activation to induce cell proliferation, survival and potential tumorigenesis. These mutations can also confer resistance to therapeutic intervention. Identifying acquired mutations, not present in normal patient tissue, in tumor derived material, including CTC’s and DTC’s, is important to diagnostic redisposition testing for cancer, prognosis and can ultimately inform targeted therapy. This application notes demonstrates a highly efficient, low-cost assay platform to detect PIK3CA somatic mutations.