Heterogeneity of Acquired Resistance to Anti-EGFR Monoclonal Antibodies in Patients with Metastatic Colorectal Cancer.

Abstract

<b>Purpose:</b> Even if <i>RAS-BRAF</i> wild-type and <i>HER2/MET</i>-negative metastatic colorectal cancer (mCRC) patients frequently respond to anti-EGFR mAbs, acquired resistance almost invariably occurs. Mechanisms of resistance to EGFR blockade include the emergence of <i>KRAS</i>, <i>NRAS</i>, and <i>EGFR</i> extracellular domain mutations as well as <i>HER2/MET</i> alterations. However, these findings derive from retrospective studies that analyzed one single resistance mechanism at a time; moreover, it is still unclear how molecular heterogeneity affects clonal evolution in patients. In this work, we aimed at extensively characterizing and correlating the molecular characteristics of tissue- and blood-based data in a prospective cohort of patients with mCRC who received anti-EGFR antibodies.<b>Experimental design:</b> Twenty-two <i>RAS</i>-<i>BRAF</i> wild-type, <i>HER2/MET</i>-negative mCRC patients progressing on anti-EGFR therapy after initial response underwent rebiopsy. Next-generation sequencing and silver <i>in situ</i> hybridization (SISH)/IHC analyses were performed both on archival tumors and postprogression samples. Circulating tumor (ctDNA) molecular profiles were obtained in matched tissue-plasma samples.<b>Results:</b><i>RAS</i> mutations and <i>HER2/MET</i> amplification were the most frequently detected resistance mechanisms in both tissue and blood sample analysis. On the other hand, <i>BRAF</i> and <i>EGFR</i> ectodomain mutations were much rarer. Patients with acquired <i>MET</i> amplification showed worse PFS on anti-EGFRs. We detected both intralesion heterogeneity, as suggested by co-occurrence of different resistance mechanisms in the same sample, and interlesion heterogeneity. The combined analysis of tissue and blood (ctDNA) results highlights the complexity of clonal evolution triggered by EGFR blockade.<b>Conclusions:</b> Our results indicate that it may be extremely challenging to target the complex landscape of molecular heterogeneity associated with emergence of resistance to targeted therapies in patients with mCRC. <i>Clin Cancer Res; 23(10); 2414-22. ©2016 AACR</i>.