The Receptor Tyrosine Kinase RON and Its Isoforms as Therapeutic Targets in Ewing Sarcoma

Abstract

The receptor tyrosine kinase (RTK) RON is linked to an aggressive metastatic phenotype of carcinomas. While gaining interest as a therapeutic target, RON remains unstudied in sarcomas. In Ewing sarcoma, we identified RON among RTKs conferring resistance to insulin-like growth factor-1 receptor (IGF1R) targeting. Therefore, we explored RON in pediatric sarcoma cell lines and an embryonic Tg(kdrl:mCherry) zebrafish model, using an shRNA-based approach. To examine RON-IGF1R crosstalk, we employed the clinical-grade monoclonal antibody IMC-RON8, alone and together with the IGF1R-antibody IMC-A12. RON silencing demonstrated functions in vitro and in vivo, particularly within micrometastatic cellular capacities. Signaling studies revealed a unidirectional IGF1-mediated cross-activation of RON. Yet, IMC-A12 failed to sensitize cells to IMC-RON8, suggesting additional mechanisms of RON activation. Here, RT-PCR revealed that childhood sarcomas express short-form RON, an isoform resistant to antibody-mediated targeting. Interestingly, in contrast to carcinomas, treatment with DNA methyltransferase inhibitor did not diminish but increased short-form RON expression. Thus, this first report supports a role for RON in the metastatic progression of Ewing sarcoma. While principal molecular functions appear transferrable between carcinomas, Ewing sarcoma and possibly more common sarcoma subtypes, RON highlights that specific regulations of cellular networks and isoforms require better understanding to successfully transfer targeting strategies.

Ewing Sarcoma—Diagnosis, Treatment, Clinical Challenges and Future Perspectives

“…The early-stage animals are optically transparent, making it possible to track metastatic cell dissemination with high resolution in a large number of test subjects. This method has been used to investigate a novel mechanism of NOTCH-induced tumor suppression involving modulation of the deacetylase SIRT1 [ 386 ]; to test the effects of the MDM2 inhibitor Nutlin-3 in combination with the small molecule YK-4-279 [ 387 ]; a role for receptor tyrosine kinase RON in EwS metastatic progression [ 388 ]; and to demonstrate synergistic effects of aurora kinase A inhibitors when combined with FAK inhibitors [ 389 ]. In one particularly notable study, Franzetti and co-workers demonstrated that plasticity in the expression level and activity of the EWSR1-FLI1 oncofusion affects the balance of proliferative states with those favoring migration, invasion and metastasis [ 285 ].…”