Abstract
The loss of function (LoF) of NF1 is the third most frequent mutation that drives hyperactivated RAS and tumor growth in >10% of melanomas. NF1LoF melanoma cells, however, do not show consistent sensitivity to individual MEK, ERK, or PI3K/mTOR inhibitors. Here, we perform a targeted kinase inhibitor screen and identify a tool compound, named MTX-216, to be highly effective in blocking NF1LoF melanoma cells. Single-cell analysis links drug-induced cytotoxicity to effective co-suppression of proliferation marker Ki-67 and the ribosomal S6 phosphorylation, an integrator of multiple RAS-mediated signaling pathways. Using a combination of kinome selectivity assay, transcriptomic analysis, and genetic experiments, we find the anti-tumor efficacy of MTX-216 to be dependent on its ability to inhibit not only PI3K (its nominal target) but also SYK, and suppression of a group of genes that regulate mitochondrial electron transport chain and whose expression is associated with poor survival in NF1LoF melanoma patients. Furthermore, combinations of inhibitors targeting either MEK or PI3K/mTOR with an independent SYK kinase inhibitor or SYK knockdown show favorable effects. These studies provide a path to exploit SYK dependency to selectively block NF1LoF melanoma cells.
Statement of significance NF1LoF melanomas represent a subtype with hyperactivated RAS signaling, for which currently no targeted therapies are clinically available. Our systems pharmacology studies identify SYK as a new vulnerability in NF1LoF melanoma cells.
Competing Interest Statement
C.W. is a cofounder of Mekanistic Therapeutics and has ownership interest in the same. J.S.-L. is a cofounder of Mekanistic Therapeutics and has ownership interest in the same. The other authors declare that they have no competing interests.
Footnotes
Financial support:
This work was supported by the Turner-McConnell Fund for Drug Discovery and the University of Michigan Rogel Cancer Center Support Grant (P30-CA046592), University of Virginia Cancer Center Support Grant (P30-CA044579), Department of Defense PRCRP Career Development Award W81XWH1810427 (to MFS), NIH grants U54-CA274499, R35-GM133404 and R01-CA249229 (to MFS), NIH grants R01-CA220199 and R01-CA241764 (to JSL), and the University of Michigan Rackham Merit Fellowship (to CA).
Disclosure of Potential Conflicts of Interest:
C.W. is a cofounder of Mekanistic Therapeutics and has ownership interest in the same. J.S.-L. is a cofounder of Mekanistic Therapeutics and has ownership interest in the same. The other authors declare that they have no competing interests.
Additional experiments and analyses were performed and presented in new Figures and Supplements.