Circulating Tumour DNA Sequencing Identifies a Genetic Resistance-Gap in Colorectal Cancers with Acquired Resistance to EGFR-Antibodies and Chemotherapy.

Abstract

Epidermal growth factor receptor antibodies (EGFR-Abs) confer a survival benefit in patients with <i>RAS</i> wild-type metastatic colorectal cancer (mCRC), but resistance invariably occurs. Previous data showed that only a minority of cancer cells harboured known genetic resistance drivers when clinical resistance to single-agent EGFR-Abs had evolved, supporting the activity of non-genetic resistance mechanisms. Here, we used error-corrected ctDNA-sequencing (ctDNA-Seq) of 40 cancer genes to identify drivers of resistance and whether a genetic resistance-gap (a lack of detectable genetic resistance mechanisms in a large fraction of the cancer cell population) also occurs in <i>RAS</i> wild-type mCRCs treated with a combination of EGFR-Abs and chemotherapy. We detected one <i>MAP2K1/MEK1</i> mutation and one <i>ERBB2</i> amplification in 2/3 patients with primary resistance and <i>KRAS, NRAS, MAP2K1/MEK1</i> mutations and <i>ERBB2</i> aberrations in 6/7 patients with acquired resistance. In vitro testing identified MAP2K1/MEK1 P124S as a novel driver of EGFR-Ab resistance. Mutation subclonality analyses confirmed a genetic resistance-gap in mCRCs treated with EGFR-Abs and chemotherapy, with only 13.42% of cancer cells harboring identifiable resistance drivers. Our results support the utility of ctDNA-Seq to guide treatment allocation for patients with resistance and the importance of investigating further non-canonical EGFR-Ab resistance mechanisms, such as microenvironmentally-mediated resistance. The detection of <i>MAP2K1</i> mutations could inform trials of MEK-inhibitors in these tumours.