Proteome-wide Profiling of Clinical PARP Inhibitors Reveals Compound-Specific Secondary Targets

Knezevic, Claire E.; Wright, Gabriëla; Rix, Lily; Kim, Woosuk; Kuenzi, Brent M.; Luo, Yunting; Watters, January M.; Koomen, John M.; Haura, Eric B.; Monteiro, Alvaro N.A.; Radu, Caius G.; Lawrence, Harshani R.; Rix, Uwe

doi:10.1016/j.chembiol.2016.10.011

Cited by 92 publications

(89 citation statements)

References 55 publications

(75 reference statements)

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“…Many effective anticancer drugs simultaneously inhibit multiple cellular targets (Knezevic et al, 2016;Wright et al, 2017). This phenomenon, termed polypharmacology, is probably common to many FDA-approved drugs and its further study offers the potential to repurpose agents that have already undergone clinical safety testing.…”

Section: Discussionmentioning

confidence: 99%

Dabrafenib inhibits the growth of BRAF‐WT cancers through CDK16 and NEK9 inhibition

et al. 2017

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Although the BRAF inhibitors dabrafenib and vemurafenib have both proven successful against BRAF‐mutant melanoma, there seem to be differences in their mechanisms of action. Here, we show that dabrafenib is more effective at inhibiting the growth of NRAS‐mutant and KRAS‐mutant cancer cell lines than vemurafenib. Using mass spectrometry‐based chemical proteomics, we identified NEK9 and CDK16 as unique targets of dabrafenib. Both NEK9 and CDK16 were highly expressed in specimens of advanced melanoma, with high expression of both proteins correlating with a worse overall survival. A role for NEK9 in the growth of NRAS‐ and KRAS‐mutant cell lines was suggested by siRNA studies in which silencing was associated with decreased proliferation, cell cycle arrest associated with increased p21 expression, inhibition of phospho‐CHK1, decreased CDK4 expression, and the initiation of a senescence response. Inhibition of CDK4 but not CHK1 recapitulated the effects of NEK9 silencing, indicating this to be the likely mechanism of growth inhibition. We next turned our attention to CDK16 and found that its knockdown inhibited the phosphorylation of the Rb protein at S780 and increased expression of p27. Both of these effects were phenocopied in NRAS‐ and KRAS‐mutant cancer cells by dabrafenib, but not vemurafenib. Combined silencing of NEK9 and CDK16 was associated with enhanced inhibition of melanoma cell proliferation. In summary, we have identified dabrafenib as a potent inhibitor of NEK9 and CDK16, and our studies suggest that inhibition of these kinases may have activity against cancers that do not harbor BRAF mutations.

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Section: Discussionmentioning

confidence: 99%

Dabrafenib inhibits the growth of BRAF‐WT cancers through CDK16 and NEK9 inhibition

et al. 2017

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“…22, 42 All enrichments were performed in duplicate. i-AZD1775 was immobilized on NHS-activated beads and blocked overnight.…”

Section: Methodsmentioning

confidence: 99%

Dual Targeting of WEE1 and PLK1 by AZD1775 Elicits Single Agent Cellular Anticancer Activity

et al. 2017

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Inhibition of the WEE1 tyrosine kinase enhances anticancer chemotherapy efficacy. Accordingly, the WEE1 inhibitor AZD1775 (previously MK-1775) is currently under evaluation in clinical trials for cancer in combination with chemotherapy. AZD1775 has been reported to display high selectivity and is therefore used in many studies as a probe to interrogate WEE1 biology. However, AZD1775 also exhibits anticancer activity as a single agent although the underlying mechanism is not fully understood. Using a chemical proteomics approach, we here describe a proteome-wide survey of AZD1775 targets in lung cancer cells and identify several previously unknown targets in addition to WEE1. In particular, we observed polo-like kinase 1 (PLK1) as a new target of AZD1775. Importantly, in vitro kinase assays showed PLK1 and WEE1 to be inhibited by AZD1775 with similar potency. Subsequent loss-of-function experiments using RNAi for WEE1 and PLK1 suggested that targeting PLK1 enhances the pro-apoptotic and antiproliferative effects observed with WEE1 knockdown. Combination of RNAi with AZD1775 treatment suggested WEE1 and PLK1 to be the most relevant targets for mediating AZD1775’s anticancer effects. Furthermore, disruption of WEE1 by CRISPR-Cas9 sensitized H322 lung cancer cells to AZD1775 to similar extent as the potent PLK1 inhibitor BI-2536 suggesting a complex crosstalk between PLK1 by WEE1. In summary, we show that AZD1775 is a potent dual WEE1 and PLK1 inhibitor, which limits its use as a specific molecular probe for WEE1. However, PLK1 inhibition makes important contributions to the single agent mechanism of action of AZD1775 and enhances its anticancer effects.

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“…Figure 3 shows one of the determinants for such selectivity – niraparib binds within the nicotinamide binding pocket in the ADP-ribosyl transferase catalytic site and also makes contacts with the regulatory subdomains of PARP1 and PARP2 (E763 & D766 in PARP1). Target maps for the four clinically successful inhibitors olaparib, veliparib, niraparib , and rucaparib , were recently developed using a mass spectroscopy based chemical proteomics assay (Knezevic et al, 2016). Despite the existence of many NAD + -binding proteins in the cell, PARP inhibitors as a class displayed high target selectivity towards PARP1, PARP2 and several of their interacting partners.…”

Section: Parp Inhibitorsmentioning

confidence: 99%

Small-Molecule Inhibitors Targeting DNA Repair and DNA Repair Deficiency in Research and Cancer Therapy

Hengel

Spies

2017

Cell Chemical Biology

120

126

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To maintain stable genomes and to avoid cancer and aging, cells need to repair a multitude of deleterious DNA lesions, which arise constantly in every cell. Processes that support genome integrity in normal cells, however, allow cancer cells to develop resistance to radiation and DNA damaging chemotherapeutics. Chemical inhibition of the key DNA repair proteins and pharmacologically induced synthetic lethality have become instrumental in both dissecting the complex DNA repair networks and as promising anticancer agents. The difficulty in capitalizing on synthetically lethal interactions in cancer cells is that many potential targets do not possess well-defined small molecule binding determinates. In this review we will discuss several successful campaigns to identify and leverage small-molecule inhibitors of the DNA repair proteins, from PARP1, a paradigm case for clinically successful small molecule inhibitors, to coveted new targets, such as RAD51 recombinase, RAD52 DNA repair protein, MRE11 nuclease and WRN DNA helicase.

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Proteome-wide Profiling of Clinical PARP Inhibitors Reveals Compound-Specific Secondary Targets

Cited by 92 publications

References 55 publications

Dabrafenib inhibits the growth of BRAF‐WT cancers through CDK16 and NEK9 inhibition

Dabrafenib inhibits the growth of BRAF‐WT cancers through CDK16 and NEK9 inhibition

Dual Targeting of WEE1 and PLK1 by AZD1775 Elicits Single Agent Cellular Anticancer Activity

Small-Molecule Inhibitors Targeting DNA Repair and DNA Repair Deficiency in Research and Cancer Therapy

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