Chemoproteomics-based kinome profiling and target deconvolution of clinical multi-kinase inhibitors in primary chronic lymphocytic leukemia cells

Kruse, Ulrich; Pallasch, Christian P.; Bantscheff, Marcus; Eberhard, Dirk; Frenzel, Laurent; Ghidelli, Sonja; Maier, S.; Werner, Thilo; Wendtner, Clemens‐Martin; Drewes, Gerard

doi:10.1038/leu.2010.233

Cited by 67 publications

(68 citation statements)

References 49 publications

(70 reference statements)

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“…However, dasatinib inhibits Src kinase activation in CLLcellsandelicitsasubstantialportionofsample-specific biologicalresponseat100-foldlowerconcentrationsthanare requiredforefficientapoptosisinductionbysorafenib.When testedataconcentrationof1mM,dasatinibandsorafenibinduced little if any apoptosis in CLL samples and were both ratedasweakinducersofapoptosisascomparedtothecyclindependent kinase (CDK) inhibitors BMS-387032 and flavopiridol [26].…”

Section: Discussionmentioning

confidence: 99%

Comparison of the Effects of Two Kinase Inhibitors, Sorafenib and Dasatinib, on Chronic Lymphocytic Leukemia Cells

et al. 2012

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Background: With sorafenib displaying the highest affinities for Flt3, VEGFR (vascular endothelial growth factor receptor) and Raf and dasatinib for Abl and Src kinases, the profiles of kinases targeted by these inhibitors differ strongly. Materials and Methods: Dose-dependent effects of the inhibitors on freshly isolated chronic lymphocytic leukemia (CLL) cells were assessed as increased phosphatidylserine exposure. Inhibition by sorafenib and dasatinib of survival and anti-apoptotic signaling in CLL cells was examined by Western blot analysis. Results: Sorafenib uniformly induced apoptosis in CLL lymphocytes with a concentration inhibiting by 50% (IC50) of 8 mM, whereas the response to dasatinib was heterogeneous with the onset of inhibition at submicromolar concentrations but with IC50 values below 25 mM in only a few samples. At the respective pharmacologically achievable plasma concentrations, the inhibitors showed more efficient apoptosis induction by sorafenib than by dasatinib and less than additive mutual enhancement in combination. Co-culture with the bone marrow stroma cell line HS-5 increased the viability of untreated CLL cells but did not protect from sorafenib-induced apoptosis. Conclusions: Sorafenib or dasatinib displayed sigmoidal or saturation-type dose-response relationships for apoptosis induction, which were uniform or highly divergent, respectively, among individual CLL samples and therefore might complement each other in their clinical potential for CLL.

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Section: Discussionmentioning

confidence: 99%

Comparison of the Effects of Two Kinase Inhibitors, Sorafenib and Dasatinib, on Chronic Lymphocytic Leukemia Cells

et al. 2012

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“…Most patients with CML were generated by Bcr-Abl, a fusion gene formed by a reciprocal chromosomal translocation t(9;22)(q34;q11) (39)(40)(41). Both patients with HES with FIP1L1-PDGFRa addiction and patients with CML generally respond to imatinib.…”

Section: Discussionmentioning

confidence: 99%

Cyclin-Dependent Kinase 7/9 Inhibitor SNS-032 Abrogates FIP1-like-1 Platelet-Derived Growth Factor Receptor α and Bcr-Abl Oncogene Addiction in Malignant Hematologic Cells

Chen

Sun

et al. 2012

Clinical Cancer Research

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Purpose: The "gate-keeper" mutations T674I platelet-derived growth factor receptor a (PDGFRa) in hypereosinophilic syndrome (HES) and T315I Bcr-Abl in chronic myeloid leukemia (CML) are resistant to imatinib and the second-generation small-molecule tyrosine kinase inhibitors (TKI). However, to combat acquired resistance to imatinib, an alternative approach is to decrease the expression of the addicted gene to efficiently kill resistant malignant hematologic cells. The purpose of this study was to evaluate the strategy of shutting down the transcription and expression of FIP1-like-1 (FIP1L1)-PDGFRa and Bcr-Abl with SNS-032, an inhibitor of cyclin-dependent kinase 7 (CDK7) and CDK9 in phase I clinical trials.Experimental Design: The effects of SNS-032 on PDGFRa and Bcr-Abl signaling pathways, apoptosis, and cell cycling were analyzed in TKI-resistant cells of HES and CML. The in vivo antitumor activity of SNS-032 was assessed with xenografted BaF3-T674I FIP1L1-PDGFRa and KBM5-T315I Bcr-Abl cells in nude mouse models.Results: SNS-032 inhibited the phosphorylation on Ser5 and Ser2 of RNA polymerase II. SNS-032 decreased both the mRNA and protein levels of FIP1L1-PDGFRa and Bcr-Abl and inhibited the proliferation of malignant cells expressing FIP1L1-PDGFRa or Bcr-Abl. It also decreased the phosphorylation of downstream molecules. It induced apoptosis by triggering both the mitochondrial pathway and the death receptor pathway.Conclusions: This CDK7/9 inhibitor potently inhibits FIP1L1-PDGFRa-positive HES cells and Bcr-Ablpositive CML cells regardless of their sensitivity to imatinib. SNS-032 may have potential in treating hematologic malignancy by abrogating oncogene addiction.

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“…Kinobeads were originally developed for selectivity profiling of small molecule kinase inhibitors (22), but we and others have also shown that kinobeads can be employed to identify kinases from human tissue including tumor cells isolated from patients (25). However, direct proteomic analysis of patient samples, although desirable, is challenging because of the often limited available sample quantity, varying ratios of tumor versus stroma versus vasculature, extent of necrosis and hypoxia, infiltration of inflammatory cells, etc., as well as significant issues with candidate validation options.…”

Section: Evaluation Of C-met As a Target In Hnscc-it Has Been Suggestmentioning

confidence: 99%

“…The second element is intensitybased label-free quantitative mass spectrometry that enables the identification and relative quantification of the purified proteins across many biological samples (24). Although the Kinobeads approach was initially developed to profile the selectivity of small molecule kinase inhibitors, it also lends itself to profiling the expression of kinases in cells or tissues (22,25). In this study, we utilized the Kinobeads approach to identify systematically kinases from HNSCC cell lines that might represent novel candidate targets for individualized therapeutic intervention and/or candidate biomarkers.…”

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confidence: 99%

Quantitative Chemical Proteomics Reveals New Potential Drug Targets in Head and Neck Cancer

Doondeea

Gholami

et al. 2011

Molecular & Cellular Proteomics

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Tumors of the head and neck represent a molecularly diverse set of human cancers, but relatively few proteins have actually been shown to drive the disease at the molecular level. To identify new targets for individualized diagnosis or therapeutic intervention, we performed a kinase centric chemical proteomics screen and quantified 146 kinases across 34 head and neck squamous cell carcinoma (HNSCC) cell lines using intensity-based labelfree mass spectrometry. Statistical analysis of the profiles revealed significant intercell line differences for 42 kinases (p < 0.05), and loss of function experiments using siRNA in high and low expressing cell lines identified kinases including EGFR, NEK9, LYN, JAK1, WEE1, and EPHA2 involved in cell survival and proliferation. EGFR inhibition by the small molecule inhibitors lapatinib, gefitinib, and erlotinib as well as siRNA led to strong reduction of viability in high but not low expressing lines, confirming EGFR as a drug target in 10 -20% of HNSCC cell lines. Similarly, high, but not low EPHA2-expressing cells showed strongly reduced viability concomitant with down-regulation of AKT and ERK signaling following EPHA2 siRNA treatment or EPHA1-Fc ligand exposure, suggesting that EPHA2 is a novel drug target in HNSCC. This notion is underscored by immunohistochemical analyses showing that high EPHA2 expression is detected in a subset of HNSCC tissues and is associated with poor prognosis. Given that the approved pan-SRC family kinase inhibitor dasatinib is also a very potent inhibitor of EPHA2, our findings may lead to new therapeutic options for HNSCC patients. Importantly, the strategy employed in this study is generic and therefore also of more general utility for the identification of novel drug targets and molecular pathway markers in tumors. This may ultimately lead to a more rational approach to individualized cancer diagnosis and therapy.

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Chemoproteomics-based kinome profiling and target deconvolution of clinical multi-kinase inhibitors in primary chronic lymphocytic leukemia cells

Cited by 67 publications

References 49 publications

Comparison of the Effects of Two Kinase Inhibitors, Sorafenib and Dasatinib, on Chronic Lymphocytic Leukemia Cells

Comparison of the Effects of Two Kinase Inhibitors, Sorafenib and Dasatinib, on Chronic Lymphocytic Leukemia Cells

Cyclin-Dependent Kinase 7/9 Inhibitor SNS-032 Abrogates FIP1-like-1 Platelet-Derived Growth Factor Receptor α and Bcr-Abl Oncogene Addiction in Malignant Hematologic Cells

Quantitative Chemical Proteomics Reveals New Potential Drug Targets in Head and Neck Cancer

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