Companion Diagnostics and Cancer Biomarkers A Comprehensive Evaluation of Biomarkers Predictive of Response to PI3K Inhibitors and of Resistance Mechanisms in Head and Neck Squamous Cell Carcinoma

The PI3K/AKT/mTOR pathway is frequently activated in head and neck squamous cell carcinoma (HNSCC), but pathway inhibition has variable efficacy. Identification of predictive biomarkers and mechan-isms of resistancewould allow selection of patientsmost likely to respond andnovel therapeutic combinations. The purpose of this study was to extend recent discoveries regarding the PI3K/AKT/mTOR pathway in HNSCCbymore broadly examining potential biomarkers of response, by examining pathway inhibitorswith a diverse range of targets, and by defining mechanisms of resistance and potential combination therapies. We used reverse-phase protein arrays (RPPA) to simultaneously evaluate expression of 195 proteins; SNP array to estimate gene copy number; and mass array to identify mutations. We examined altered signaling at baseline and after pathway inhibition. Likewise, we examined the activation of the PI3K/AKT/mTOR pathway in HNSCC tumors by RPPA. Cell lines with PIK3CA mutations were sensitive to pathway inhibitors, whereas amplification status did not predict sensitivity.Whilewe identified a set of individual candidate biomarkers of response topathway inhibitors, proteomic pathway scoresdidnot correlatewith amplification ormutation and did not predict response. Several receptor tyrosine kinases, includingEGFRandERK,were activated following PI3K inhibition in resistant cells; dual pathway inhibition of PI3K and EGFR or MEK demonstrated synergy. Combined MEK and PI3K inhibition was markedly synergistic in HRAS-mutant cell lines. Our findings indicate that clinical trials of single-agent PI3K/AKT/mTORpathway inhibitors in selectedpopulations andof PI3K/EGFR or PI3K/MEK inhibitor combinations are warranted; we plan to conduct such trials.Mol Cancer Ther; 13(11); 2738–50. 2014 AACR