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A novel protein isoform of the RON tyrosine kinase receptor transforms human pancreatic duct epithelial cells

Oncogene volume 35, pages 3249–3259 (2016)Cite this article

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Abstract

The MST1R gene is overexpressed in pancreatic cancer producing elevated levels of the RON tyrosine kinase receptor protein. While mutations in MST1R are rare, alternative splice variants have been previously reported in epithelial cancers. We report the discovery of a novel RON isoform discovered in human pancreatic cancer. Partial splicing of exons 5 and 6 (P5P6) produces a RON isoform that lacks the first extracellular immunoglobulin-plexin-transcription domain. The splice variant is detected in 73% of xenografts derived from pancreatic adenocarcinoma patients and 71% of pancreatic cancer cell lines. Peptides specific to RON P5P6 detected in human pancreatic cancer specimens by mass spectrometry confirm translation of the protein isoform. The P5P6 isoform is found to be constitutively phosphorylated, present in the cytoplasm, and it traffics to the plasma membrane. Expression of P5P6 in immortalized human pancreatic duct epithelial (HPDE) cells activates downstream AKT, and in human pancreatic epithelial nestin-expressing cells, activates both the AKT and MAPK pathways. Inhibiting RON P5P6 in HPDE cells using a small molecule inhibitor BMS-777607 blocked constitutive activation and decreased AKT signaling. P5P6 transforms NIH3T3 cells and induces tumorigenicity in HPDE cells. Resultant HPDE-P5P6 tumors develop a dense stromal compartment similar to that seen in pancreatic cancer. In summary, we have identified a novel and constitutively active isoform of the RON tyrosine kinase receptor that has transforming activity and is expressed in human pancreatic cancer. These findings provide additional insight into the biology of the RON receptor in pancreatic cancer and are clinically relevant to the study of RON as a potential therapeutic target.

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Acknowledgements

We gratefully acknowledge the generosity of our patients who supported this work through both monetary contributions and/or by allowing tumor tissue to be used for research. Without their contributions, this work would not have been possible. We thank the Tsao and Klemke labs for providing HPDE and HPNE ells, respectively. Research reported in this publication was supported by the National Cancer Institute of the National Institutes of Health under Award Number R01 CA155620 (AML) and T32 CA121938 (JC). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. DNA sequencing was performed by the DNA Sequencing Shared Resource, UCSD Moore’s Cancer Center, which is funded in part by NCI Cancer Center Support Grant # 2 P30 CA023100-23.

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  1. Department of Surgery, Division of Surgical Oncology, Moores Cancer Center, University of California, San Diego, La Jolla, CA, USA

    J Chakedis, R French, M Babicky, D Jaquish, H Howard, E Mose, R Lam, P Holman, J Miyamoto, Z Walterscheid & A M Lowy

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Correspondence to A M Lowy.

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Chakedis, J., French, R., Babicky, M. et al. A novel protein isoform of the RON tyrosine kinase receptor transforms human pancreatic duct epithelial cells. Oncogene 35, 3249–3259 (2016). https://doi.org/10.1038/onc.2015.384

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