Dataset Information

Chemical phosphoproteomics reveals kinase network topologies associated to genotypes and phenotypes of cancer cells.

ABSTRACT: We compare the in vitro specificity profiles of several kinase inhibitors with their effects on cellular phosphoproteomes. For this aim, we developed an algorithm which utilizes information on kinase inhibitor selectivity to determine the most probable kinase(s) acting upstream of phosphorylation sites present in phosphoproteomics data obtained from cancer cells treated with kinase inhibitors.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Pedro Cutillas

PROVIDER: MSV000090571 | MassIVE | Sat Oct 22 21:47:00 BST 2022

SECONDARY ACCESSION(S): PXD015943

REPOSITORIES: MassIVE

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Publications

Reconstructing kinase network topologies from phosphoproteomics data reveals cancer-associated rewiring.

Hijazi Maruan M Smith Ryan R Rajeeve Vinothini V Bessant Conrad C Cutillas Pedro R PR

Nature biotechnology 20200120 4

Understanding how oncogenic mutations rewire regulatory-protein networks is important for rationalizing the mechanisms of oncogenesis and for individualizing anticancer treatments. We report a chemical phosphoproteomics method to elucidate the topology of kinase-signaling networks in mammalian cells. We identified >6,000 protein phosphorylation sites that can be used to infer >1,500 kinase-kinase interactions and devised algorithms that can reconstruct kinase network topologies from these phosph ...[more]

PMID: 31959955

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Project description:There is an unmet need for chemical tools to explore the role of the Mediator complex in human pathologies ranging from cancer to cardiovascular disease. Here we determine that CCT251545 a WNT-pathway inhibitor discovered by cell-based screening is a potent and selective chemical probe for the Mediator complex-associated protein kinases CDK8 and CDK19. CCT251545 is an ATP competitive inhibitor with >100-fold selectivity over 291 other kinases. X-ray crystallography demonstrates Type 1 binding involving insertion of the CDK8 C-terminus into the ligand binding site. In contrast to Type II inhibitors of CDK8/19 CCT251545 displays potent cell-based activity. We show that CCT251545 and close analogues not only alter WNT-pathway regulated gene expression but also other CDK8/19 targets including STAT1-regulated gene expression. Consistent with this we find that phospho-STAT1SER727 is a biomarker of CDK8 kinase activity in vitro and in vivo. Finally we demonstrate in vivo activity of CCT251545 in WNT-dependent tumours. Compound 1,2, 6 and 9 are close analogues of a 3,4,5-trisubstituted pyridine series identified from a high-throughput cell-based reporter assay of WNT signalling. They were shown to be potent and selective inhibitors of the Mediator complex-associated protein kinases CDK8 and CDK19. They are ATP competitive inhibitors with >100-fold selectivity over 291 other kinases. X-ray crystallography demonstrates Type 1 binding involving insertion of the CDK8 C-terminus into the ligand-binding site. A 3D culture model of murine intestinal-derived organoids expressing a transgenic doxycycline-inducible mutant beta-catenin expression following removal of doxycycline results in reduced WNT signalling and was compared to treatment for 24 hours with coumpound 2, 6 and 9. Compound 2 (n=3), compound 6 (n=4), compound 9 (n=3), doxycycline removal (n=4), doxycycline treatment (n=7). Samples were hybridized agianst mouse reference. Compounds 2, 6 and 9 are close analogues of a 3,4,5-trisubstituted pyridine series identified from a high-throughput cell-based reporter assay of WNT signalling. They were shown to be potent and selective inhibitors of the Mediator complex-associated protein kinases CDK8 and CDK19. They are ATP competitive inhibitors with >100-fold selectivity over 291 other kinases. X-ray crystallography demonstrates Type 1 binding involving insertion of the CDK8 C-terminus into the ligand-binding site.

Dataset Information

Chemical phosphoproteomics reveals kinase network topologies associated to genotypes and phenotypes of cancer cells.

Publications

Reconstructing kinase network topologies from phosphoproteomics data reveals cancer-associated rewiring.

Similar Datasets

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