Covalent inhibitors of EGFR family protein kinases induce degradation of human Tribbles 2 (TRIB2) pseudokinase in cancer cells

被引:58
作者
Foulkes, Daniel M. [1 ]
Byrne, Dominic P. [1 ]
Yeung, Wayland [2 ,3 ]
Shrestha, Safal [2 ,3 ]
Bailey, Fiona P. [1 ]
Ferries, Samantha [1 ,4 ]
Eyers, Claire E. [1 ,4 ]
Keeshan, Karen [5 ]
Wells, Carrow [6 ]
Drewry, David H. [6 ]
Zuercher, William J. [6 ,7 ]
Kannan, Natarajan [2 ,3 ]
Eyers, Patrick A. [1 ]
机构
[1] Univ Liverpool, Inst Integrat Biol, Dept Biochem, Liverpool L69 7ZB, Merseyside, England
[2] Univ Georgia, Inst Bioinformat, Athens, GA 30602 USA
[3] Univ Georgia, Dept Biochem & Mol Biol, Athens, GA 30602 USA
[4] Univ Liverpool, Ctr Proteome Res, Inst Integrat Biol, Liverpool L69 7ZB, Merseyside, England
[5] Univ Glasgow, Paul OGorman Leukaemia Res Ctr, Inst Canc Sci, Glasgow, Lanark, Scotland
[6] Univ North Carolina Chapel Hill, UNC Eshelman Sch Pharm, Struct Genom Consortium, Chapel Hill, NC 27599 USA
[7] Univ North Carolina Chapel Hill, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USA
基金
英国生物技术与生命科学研究理事会; 巴西圣保罗研究基金会; 加拿大创新基金会; 英国惠康基金;
关键词
NONCATALYTIC FUNCTIONS; STRUCTURAL-ANALYSIS; RESISTANT AURORA; C/EBP-ALPHA; BIBW; 2992; BINDING; TARGET; DOMAIN; ACTIVATION; DISCOVERY;
D O I
10.1126/scisignal.aat7951
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A major challenge associated with biochemical and cellular analysis of pseudokinases is a lack of target-validated small-molecule compounds with which to probe function. Tribbles 2 (TRIB2) is a cancer-associated pseudokinase with a diverse interactome, including the canonical AKT signaling module. There is substantial evidence that human TRIB2 promotes survival and drug resistance in solid tumors and blood cancers and therefore is of interest as a therapeutic target. The unusual TRIB2 pseudokinase domain contains a unique cysteine-rich C-helix and interacts with a conserved peptide motif in its own carboxyl-terminal tail, which also supports its interaction with E3 ubiquitin ligases. We found that TRIB2 is a target of previously described small-molecule protein kinase inhibitors, which were originally designed to inhibit the canonical kinase domains of epidermal growth factor receptor tyrosine kinase family members. Using a thermal shift assay, we discovered TRIB2-binding compounds within the Published Kinase Inhibitor Set (PKIS) and used a drug repurposing approach to classify compounds that either stabilized or destabilized TRIB2 in vitro. TRIB2 destabilizing agents, including the covalent drug afatinib, led to rapid TRIB2 degradation in human AML cancer cells, eliciting tractable effects on signaling and survival. Our data reveal new drug leads for the development of TRIB2-degrading compounds, which will also be invaluable for unraveling the cellular mechanisms of TRIB2-based signaling. Our study highlights that small molecule-induced protein down-regulation through drug "off-targets" might be relevant for other inhibitors that serendipitously target pseudokinases.
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页数:14
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