Abstract 191: LMTK3 is a novel regulator of oncogenic KIT in KIT-mutant cancers

Abstract

Abstract Multiple cancers, such as gastrointestinal stromal tumors (GIST) and melanoma, have been shown to be caused by somatic activating mutations in the receptor tyrosine kinase KIT. The major cause of death in patients with advanced KIT-mutant cancers is due to KIT tyrosine kinase inhibitor-resistant metastatic disease. Resistance develops almost exclusively because of secondary mutations within KIT, highlighting the importance of KIT in the proliferation and survival of these tumors. Therefore, strategies that affect KIT protein expression and maturation may be therapeutic. In order to identify regulators of KIT that may represent novel targets, we performed an siRNA screen in multiple KIT-mutant cancer cell lines. From these screens we identified lemur tyrosine kinase 3 (LMTK3) as a novel KIT regulator and potential target in KIT-mutant cancers. LMTK3 has been implicated as an important protein in multiple cancers, including leukemia, breast, and colon cancer. However, the oncogenic function of LMTK3 is not entirely clear. Despite its name, lemur tyrosine kinase 3 is, in fact, a serine/ threonine kinase. Beyond its enzymatic kinase function, LMTK3 has also been shown to have important scaffolding functions. LMTK3 has not been previously known to interact with KIT, but does regulate the stability of other receptors. We have found that LMTK3 silencing reduced the viability of all KIT-mutant GIST and melanoma cells tested to date, including those with ATP binding pocket and activation loop resistance mutations. Importantly, LMTK3 silencing decreased the viability of KIT-mutant cells specifically, as silencing of LMTK3 in cells lacking KIT mutations did not affect their viability. Further, we found the decrease in cell viability was due to induction of apoptosis, as seen by cleavage of PARP within 96 hours of LMTK3 silencing. Because these cells depend so heavily on KIT and loss of KIT signaling results in cell death, we hypothesized that LMTK3 silencing may be affecting this pathway. Indeed, with LMTK3 silencing there was a significant decrease in phosphorylated KIT (Y719), which was accompanied by decreased phospho-AKT (S473). We also observed a robust decrease in total KIT protein levels. However, there was no decrease in KIT transcript levels, suggesting that LMTK3 plays a role in regulating KIT at the protein level. While other KIT protein regulators, such as HSP90, have been suggested as therapeutic targets for GIST, the ubiquitous functions of these targets makes these treatments sub-optimal, often causing toxicity. Silencing of LMTK3, on the other hand, specifically kills KIT-mutant cancer cells. Furthermore, as a kinase, inhibition of LMTK3 by a small molecule may be possible, and we are in the process of identifying LMTK3 inhibitors. LMTK3 is a positive regulator of oncogenic KIT in GIST and melanoma and represents a novel, tractable target in KIT-mutant cancers. Citation Format: Lillian R. Klug, Jeffery W. Tyner, Michael C. Heinrich. LMTK3 is a novel regulator of oncogenic KIT in KIT-mutant cancers. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 191.