Aims
Malignant pleural mesothelioma (MPM) is an aggressive inflammatory cancer. Using a phospho-receptor tyrosine kinase array, we identified RON as frequently activated in MPM patient samples and cell lines. RON is a member of the MET proto-oncogene family and is bound by macrophage stimulating protein (MSP). The aim of this study was to examine the MSP-RON signalling axis in MPM.
Methods
Immunohistochemistry was performed for RON, MSP and CD68 on a MPM tissue-microarray (n=132). MSP levels were determined in MPM and control serum samples (n=40). Expression levels were correlated with clinico-pathological data. MPM cell lines and a normal mesothelial cell line were screened for the expression of RON and MSP at the protein (Western) and mRNA (RT-PCR) level. Downstream mediators affected by MSP stimulation were identified using a proteome profiler array. The effect of MSP, IMC-RON8 (humanised IgG1 monoclonal antibody), LCRF004 (small molecule inhibitor) and NRWHE (small peptide) was examined using proliferation (BrdU ELISA), viability and migration (xCELLigence) assays. An xenograft study was completed with LCRF004.
Results
High positivity for total RON by IHC was an independent predictor of favourable prognosis. Additionally, elevated expression levels of MSP correlated with better survival. The protein and mRNA levels of RON and MSP were differentially expressed in MPM cell lines. MSP treatment resulted in the phosphorylation of a number of kinases including Src. Treatment with LCRF004 resulted in a significant decrease in proliferation, viability, migration and reduced tumour growth in vivo (p<0.05, compared with vehicle control).
Conclusion
The seemingly counter intuitive results obtained from TMA studies and cell line data, may be dependant on RON isoform expression. Nevertheless, the in vivo and in vitro data generated in this study, indicates that the MSP-RON signalling axis is a potential target in MPM. LCRF004 is an encouraging novel targeted therapeutic agent in this disease.