Spatio-temporal tumor heterogeneity in metastatic CRC tumors: a mutational-based approach

// Sofia del Carmen 1, * , Jose Maria Sayagues 2, * , Oscar Bengoechea 1 , Maria Fernanda Anduaga 3 , Jose Antonio Alcazar 3 , Ruth Gervas 1 , Jacinto Garcia 3 , Alberto Orfao 2, ** , Luis Munoz Bellvis 3, ** , Maria Eugenia Sarasquete 4, ** and Maria del Mar Abad 1, ** 1 Department of Pathology and IBSAL, University Hospital of Salamanca, Salamanca, Spain 2 Cytometry Service-NUCLEUS, Department of Medicine, Cancer Research Center (IBMCC-CSIC/USAL) and IBSAL, University Hospital of Salamanca, Salamanca, Spain 3 General and Gastrointestinal Surgery Service and IBSAL, University Hospital of Salamanca, Salamanca, Spain 4 Hematology Service, University Hospital of Salamanca, Salamanca, Spain * These authors have contributed equally to this work ** These authors have contributed equally to this work and they should be considered as senior last authors and corresponding authors Correspondence to: Maria Eugenia Sarasquete, email: mealonsos@saludcastillayleon.es Maria del Mar Abad, email: marabad@usal.es Keywords: colorectal cancer; clonal evolution; anti-EGFR; mutational profile; tumor heterogeneity Received: June 30, 2018      Accepted: August 10, 2018      Published: September 28, 2018 ABSTRACT It is well known that activating mutations in the KRAS and NRAS genes are associated with poor response to anti-EGFR therapies in patients with metastatic colorectal cancer (mCRC). Approximately half of the patients with wild-type (WT) KRAS colorectal carcinoma do not respond to these therapies. This could be because the treatment decision is determined by the mutational profile of the primary tumor, regardless of the presence of small tumor subclones harboring RAS mutations in lymph nodes or liver metastases. We analyzed the mutational profile of the KRAS , NRAS , BRAF and PI3KCA genes using low-density microarray technology in samples of 26 paired primary tumors, 16 lymph nodes and 34 liver metastases from 26 untreated mCRC patients (n=76 samples). The most frequent mutations found in primary tumors were KRAS (15%) and PI3KCA (15%), followed by NRAS (8%) and BRAF (4%). The distribution of the mutations in the 16 lymph node metastases analyzed was as follows: 4 (25%) in KRAS gene, 3 (19%) in NRAS gene and 1 mutation each in PI3KCA and BRAF genes (6%). As expected, the most prevalent mutation in liver metastasis was in the KRAS gene (35%), followed by PI3KCA (9%) and BRAF (6%). Of the 26 cases studied, 15 (58%) displayed an overall concordance in the mutation status detected in the lymph node metastases and liver metastases compared with primary tumor, suggesting no clonal evolution. In contrast, the mutation profiles differed in the primary tumor and lymph node/metastases samples of the remaining 11 patients (48%), suggesting a spatial and temporal clonal evolution. We confirm the presence of different mutational profiles among primary tumors, lymph node metastases and liver metastases. Our results suggest the need to perform mutational analysis in all available tumor samples of patients before deciding to commence anti-EGFR treatment.