Abstract 2079: EpHA2 is an essential driver of invasion and a novel target in KRAS mutant colorectal cancer
Authors: Dunne Philip D. , McArt Darragh G. , Blayney Jaine K. , Dasgupta Sonali , Salto-Tellez Manuel , Johnston Patrick G. , Schaeybroeck Sandra Van 
Published: May 2015

Background

Colorectal cancer (CRC) is the third most common cancer worldwide. Among patients with early stage CRC, there are subsets of patients who will not benefit from adjuvant 5-FU or 5-FU/oxaliplatin treatment, highlighting the need for improved therapy options for early stage disease. In addition mutations in the KRAS proto-oncogene are found in 40%-45% of CRC patients and this mutation predicts resistance to EGFR mAb therapies. More recently it has been shown that treatments, such as bevacizumab or cetuximab, which prolong survival in the advanced disease setting, cannot be translated into successful adjuvant treatment strategies.

The aim of this study was to identify therapy options which will be effective in the adjuvant setting and also whose knock-down is synthetically lethal with KRAS mutation in CRC models.

Method

A novel systems biology approach combined profiling of primary KRASMT/WT CRC tissues and isogenic paired KRASMT/WT CRC xenografts models. Metacore pathway analysis identified individual genes from novel KRAS-dependent pathways for incorporation into a primary RNAi screen. This screen was crossed with receptor tyrosine kinase analysis of our previously published pre-clinical adjuvant invasive cell line models. Our findings were validated by in vitro screening and expression analysis in CRC FFPE sections was measured using IHC.

Results

Our comprehensive approach identified 30 signalling pathways and 160 genes associated with KRASMT CRC cancer. Our siRNA screen confirmed a number of novel KRAS synthetic lethal target genes with a critical role in EMT and cell adhesion/migration. Down-regulation of members of the Eph/Ephrin family, in particular EpHA2, was found to be lethal in KRASMT but not KRASWT cells. This lethality was further synergistically enhanced in the presence of chemotherapy treatment.

Overexpression of EphA2 was also identified in each of our invasive isogenic cell line models, regardless of genetic background, and EpHA2 silencing inhibited migration and invasion in a panel of CRC cells. EpHA2 expression has previously been reported as an essential component in cancer cells's ability to form tumors in vivo and in-line with this our EpHA2 overexpressing invasive subpopulations displayed an increased ability to form colonies. Importantly, high EpHA2 mRNA and protein expression were found to be associated with poor overall survival in both early stage CRC tissues and in advanced disease.

Conclusion

Using our novel comprehensive systems biology approach we have identified novel KRAS synthetic lethal targets and pathways. Combining these findings with targets which are effective against our pre-clinical adjuvant models we have identified EpHA2 as a key driver of invasion and migration and a synthetically lethal target in KRASMT CRC. In addition, we show that EpHA2 is a poor prognostic marker and an important novel target for KRASMT CRC tumors in both the adjuvant and advanced disease setting.

Citation Format: Philip D. Dunne, Darragh G. McArt, Jaine K. Blayney, Sonali Dasgupta, Manuel Salto-Tellez, Patrick G. Johnston, Sandra Van Schaeybroeck. EpHA2 is an essential driver of invasion and a novel target in KRAS mutant colorectal cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2079. doi:10.1158/1538-7445.AM2014-2079