Survey of Tyrosine Kinase Signaling Reveals ROS Kinase Fusions in Human Cholangiocarcinoma

Abstract

Cholangiocarcinoma, also known as bile duct cancer, is the second most common primary hepatic carcinoma with a median survival of less than 2 years. The molecular mechanisms underlying the development of this disease are not clear. To survey activated tyrosine kinases signaling in cholangiocarcinoma, we employed immunoaffinity profiling coupled to mass spectrometry and identified DDR1, EPHA2, EGFR, and ROS tyrosine kinases, along with over 1,000 tyrosine phosphorylation sites from about 750 different proteins in primary cholangiocarcinoma patients. Furthermore, we confirmed the presence of ROS kinase fusions in 8.7% (2 out of 23) of cholangiocarcinoma patients. Expression of the ROS fusions in 3T3 cells confers transforming ability both in vitro and in vivo, and is responsive to its kinase inhibitor. Our data demonstrate that ROS kinase is a promising candidate for a therapeutic target and for a diagnostic molecular marker in cholangiocarcinoma. The identification of ROS tyrosine kinase fusions in cholangiocarcinoma, along with the presence of other ROS kinase fusions in lung cancer and glioblastoma, suggests that a more broadly based screen for activated ROS kinase in cancer is warranted.

Immunohistochemical detection of ROS1 is useful for identifying ROS1 rearrangements in lung cancers

“…We further suspect that ROS1 immunohistochemistry may find additional utility in wider clinical field in the future because ROS1 rearrangements have also been reported in a growing number of nonpulmonary tumors. 19,27,28 …”

Oxido-reductive regulation of vascular remodeling by receptor tyrosine kinase ROS1

“…As cell adhesion is critical to the pathogenesis of vascular remodeling, the structural properties of ROS1 make it attractive as a candidate for a role in such processes. From a clinical standpoint, ROS1 has been described predominantly in the setting of chromosomal translocations creating gene fusions in highly malignant cancers (33)(34)(35)(36)(37)(38). ROS1 is an attractive therapeutic target, not only because it drives proliferation, but also because of its preferential expression.…”

Inhibitory effect of SPE‐39 due to tyrosine phosphorylation and ubiquitination on the function of Vps33B in the EGF‐stimulated cells

“…An anti-SPE-39 rabbit polyclonal antibody was raised against peptides having an additional cysteine residue at the N-terminal end of the SPE-39 amino acid sequence, KGDEEEYWNSSKF (amino acids [5][6][7][8][9][10][11][12][13][14][15][16][17]. The peptides were coupled to keyhole limpet hemocyanin and used to immunize rabbits.…”

“…Tyr-11 was identified as the site of phosphorylation in fibroblast growth factor-3 (FGF3)-stimulated cells, as well as in pervanadate-and calyculin-treated T-cell leukemia cells [9][10][11].…”

“…However, the precise mechanisms by which this occurs remain obscured. SPE-39 (also known as FLJ12707 or C14ORF133) was originally identified through tyrosine phosphoproteomics, and it has been reported that Tyr-11 is phosphorylated following certain stimuli in cultured cells [9][10][11]. However, further characteristics of its tyrosine phosphorylation have yet to be conducted.…”