Kinome Profiling Identifies MARK3 and STK10 As Potential Therapeutic Targets in Uveal Melanoma

Authors

Usman Baqai
Alison M Kurimchak
Isabella V Trachtenberg
Timothy J Purwin
Jelan I Haj
Anna Han
Kristine Luo
Nikole Fandino Pachon
Angela Jeon
Vivian Chua
Michael A Davies
J Silvio Gutkind
Jeffrey L Benovic
James S Duncan
Andrew E Aplin

Publication Date

12-1-2023

Journal

Journal of Biological Chemistry

Abstract

Most uveal melanoma cases harbor activating mutations in either GNAQ or GNA11. Despite activation of the mitogen-activated protein kinase (MAPK) signaling pathway downstream of Gαq/11, there are no effective targeted kinase therapies for metastatic uveal melanoma. The human genome encodes numerous understudied kinases, also called the "dark kinome". Identifying additional kinases regulated by Gαq/11 may uncover novel therapeutic targets for uveal melanoma. In this study, we treated GNAQ-mutant uveal melanoma cell lines with a Gαq/11 inhibitor, YM-254890, and conducted a kinase signaling proteomic screen using multiplexed-kinase inhibitors followed by mass spectrometry. We observed downregulated expression and/or activity of 22 kinases. A custom siRNA screen targeting these kinases demonstrated that knockdown of microtubule affinity regulating kinase 3 (MARK3) and serine/threonine kinase 10 (STK10) significantly reduced uveal melanoma cell growth and decreased expression of cell cycle proteins. Additionally, knockdown of MARK3 but not STK10 decreased ERK1/2 phosphorylation. Analysis of RNA-sequencing and proteomic data showed that Gαq signaling regulates STK10 expression and MARK3 activity. Our findings suggest an involvement of STK10 and MARK3 in the Gαq/11 oncogenic pathway and prompt further investigation into the specific roles and targeting potential of these kinases in uveal melanoma.

Keywords

Humans, Cell Line, Tumor, GTP-Binding Protein alpha Subunits, Gq-G11, Melanoma, Mutation, Protein Serine-Threonine Kinases, Proteomics, Uveal Neoplasms, Uveal Melanoma, cancer, microtubule affinity-regulating kinase 3, melanoma, g protein, mitogen-activated protein kinase (MAPK), guanine nucleotide-binding protein G(q) subunit alpha, proteomics, serine/threonine-protein kinase 10

DOI

10.1016/j.jbc.2023.105418

PMID

37923138

PMCID

PMC10716579

PubMedCentral® Posted Date

11-3-2023

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes