RT Journal Article
SR Electronic
T1 Kinome Profiling of Gastrointestinal Stromal Tumors Using Multiplexed Inhibitor Beads and Mass Spectrometry Identifies Wee1 as Candidate Therapeutic Target
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.07.138693
DO 10.1101/2020.06.07.138693
A1 Shuai Ye
A1 Dinara Sharipova
A1 Marya Kozinova
A1 Lilli Klug
A1 Jimson D’Souza
A1 Martin G. Belinsky
A1 Katherine J. Johnson
A1 Margret B. Einarson
A1 Kathy Cai
A1 Karthik Devarajan
A1 Yan Zhou
A1 Samuel Litwin
A1 Michael C. Heinrich
A1 Ronald DeMatteo
A1 Margaret von Mehren
A1 James Duncan
A1 Lori Rink
YR 2020
UL http://biorxiv.org/content/early/2020/06/08/2020.06.07.138693.abstract
AB Gastrointestinal stromal tumor (GIST) management has been revolutionized by the identification of activating mutations in KIT and PDGFRA and clinical application of receptor tyrosine kinase (RTK) inhibitors in advanced disease. Stratification of GIST into molecularly defined subsets provides insight into clinical behavior and response to approved targeted therapies. Although these RTK inhibitors are effective in most GIST, resistance remains a significant clinical obstacle. Development of effective strategies for refractory GIST requires identification of novel targets to provide additional therapeutic options. Global kinome profiling has potential to identify critical signaling networks and reveal protein kinases that are essential in GIST. Using Multiplexed Inhibitor Beads and Mass Spectrometry paired with a super-SILAC kinome standard, we explored the majority of the kinome in GIST specimens from the three most common molecular subtypes to identify novel kinase targets. Kinome profiling revealed distinct signatures in GIST subtypes. PDGFRA-mutant GIST had elevated tumor associated macrophage (TAM) kinases and immunohistochemical analysis confirmed increased TAMs present in these tumors. Kinome profiling with loss-of-function assays revealed a significant role for G2-M tyrosine kinase, Wee1, in GIST survival. In vitro and in vivo studies revealed significant efficacy of MK-1775 (Wee1 inhibitor) in combination with avapritinib in KIT and PDGFRA-mutant GIST cell lines, and notable efficacy of MK-1775 as a monotherapy in the PDGFRA-mutant line. These studies provide strong preclinical justification for the use of MK-1775 in GIST.Competing Interest StatementThe authors have declared no competing interest.