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

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.

Keywords: anti-EGFR; clonal evolution; colorectal cancer; mutational profile; tumor heterogeneity.

Figure 1. Intratumoral and intertumoral mutation heterogeneity of primary colorectal carcinomas as determined by the presence of different mutation status for KRAS, NRAS, PIK3 and BRAF genes

Only those patients with mutation heterogeneity across the three tumor samples in one or more of the four examined genes are shown (n=11). A detailed description of all patients (n=26) and samples (number and type) analyzed can be found in Table 1. Each circle represents a tumor cell clone detected in the sample analyzed. Tissue origin in represented by the following pictures: for the primary tumor; lymph node; liver metastasis. If more than one sample per tissue was studied it is mentioned in the figure (n= number of samples).