Over 15,000 Australians are diagnosed with colorectal cancer (CRC) annually. Despite available treatments, the 5-year survival rate for metastatic CRC is <14%.
Hyper-activation of the ERK/MAPK-pathway is a key driver of CRC. Genetic alterations of ERK/MAPK-pathway components (RAS, BRAF, EGFR, HER2) that drive constitutive pathway activation occur in ~60% of CRC’s, for which targeted therapies are now clinically utilised. While the remaining ~40% of CRC’s lack genetic alterations in known ERK/MAPK activators, a significant proportion still likely depend on this pathway for growth. The mechanisms driving ERK/MAPK-pathway activation in these wild-type CRC’s remain unclear, and uncovering novel drivers could enable development of new targeted therapies.
As the primary effector of the ERK/MAPK-pathway, ERK is regulated by the map-kinase phosphatase (MKP) subset of dual-specificity phosphatases (DUSP), and the role of their deletion in CRC remains unclear. We examined the deletion frequency of MKP’s in the TCGA cohort of 231 microsatellite-stable CRC’s, and identified DUSP4 as the most frequently deleted. We therefore hypothesised that DUSP4 copy loss may be an alternate mediator of ERK/MAPK activation, and an unknown driver of CRC.
To investigate this, we examined the frequency of DUSP4 deletion and the relationship with genomic alterations in other components of the ERK/MAPK-pathway (TCGA dataset) in CRCs. We found that DUSP4 copy loss occurs in ~48% of CRCs, and is significantly enriched in tumours lacking BRAF/RAS mutations (P<0.05).
Next, to determine the functional significance of DUSP4 loss in this context, we re-expressed DUSP4 in BRAF/RAS wild-type CRC cell lines with genetic deletion of DUSP4 and lacking endogenous expression. DUSP4 re-expression suppressed ERK/MAPK-pathway output, both basally and upon EGF-stimulation, consistent with its role as a critical negative regulator of ERK output. DUSP4 re-expression also significantly reduced proliferation and colony formation in BRAF/RAS WT CRC cells. Finally, consistent with its tumour suppressive effect, DUSP4 re-expressing cells were selected against during cell line expansion.
Our preliminary results support genomic DUSP4 loss as a facilitator of ERK/MAPK-pathway activation and proliferation in CRC. Validation of these findings in a mouse model is underway to determine the impact of Dusp4 deletion on colon homeostasis and Apc-driven colon tumorigenesis.