Comprehensive assay of kinase catalytic activity reveals features of kinase inhibitor selectivity

Anastassiadis, Theonie; Deacon, Sean; Devarajan, Karthik; Ma, Haiching; Peterson, Jeffrey R.

doi:10.1038/nbt.2017

Cited by 775 publications

(1,016 citation statements)

References 35 publications

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“…with RNAi knockdown) is that it provides the opportunity to easily screen transiently transfected cells. The limitation of this approach arises from off-target activity and cell toxicity exhibited by many kinase inhibitors (39). Several lines of evidence argue against Table 2.…”

Section: Discussionmentioning

confidence: 99%

The Fibroblast Growth Factor 14·Voltage-gated Sodium Channel Complex Is a New Target of Glycogen Synthase Kinase 3 (GSK3)

Shavkunov

Wildburger²,

Nenov³

et al. 2013

Journal of Biological Chemistry

160

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Section: Discussionmentioning

confidence: 99%

The Fibroblast Growth Factor 14·Voltage-gated Sodium Channel Complex Is a New Target of Glycogen Synthase Kinase 3 (GSK3)

Shavkunov

Wildburger²,

Nenov³

et al. 2013

Journal of Biological Chemistry

160

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“…To test if MAP3K9 could directly phosphorylate MEK in vitro, we performed a kinase assay and observed that purified MAP3K9 kinase domain can phosphorylate kinase-dead MEK1 (Fig. 5C) (31). To determine if the E179K mutation is a GOF mutation, we expressed MAP3K9 E179K in HEK293T cells along with a WT and kinase-dead MAP3K9 (D294A).…”

Section: Gof Mutation In Map3k9 Leads To Preferential Activation Of Tmentioning

confidence: 99%

Targeted genetic dependency screen facilitates identification of actionable mutations in FGFR4, MAP3K9, and PAK5 in lung cancer

Fawdar¹,

Trotter²,

Li³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Approximately 70% of patients with non-small-cell lung cancer present with late-stage disease and have limited treatment options, so there is a pressing need to develop efficacious targeted therapies for these patients. This remains a major challenge as the underlying genetic causes of ∼50% of non-small-cell lung cancers remain unknown. Here we demonstrate that a targeted genetic dependency screen is an efficient approach to identify somatic cancer alterations that are functionally important. By using this approach, we have identified three kinases with gain-of-function mutations in lung cancer, namely FGFR4, MAP3K9, and PAK5. Mutations in these kinases are activating toward the ERK pathway, and targeted depletion of the mutated kinases inhibits proliferation, suppresses constitutive activation of downstream signaling pathways, and results in specific killing of the lung cancer cells. Genomic profiling of patients with lung cancer is ushering in an era of personalized medicine; however, lack of actionable mutations presents a significant hurdle. Our study indicates that targeted genetic dependency screens will be an effective strategy to elucidate somatic variants that are essential for lung cancer cell viability.driver mutations | MAPK pathway | signal transduction | siRNA screen

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“…Ruxolitinib is approved for myelofibrosis and is being explored for polycythemia vera (15)(16)(17). The therapeutic efficacy of any drug is also directly related to its specific target portfolio, and kinase inhibitors are well established to have diverse selectivities across the kinome (18). To fully understand the activity of each JAK inhibitor, we performed an in-depth kinome activity profile to provide a contextual understanding of selective and nonselective antiproliferative actions of each drug.…”

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confidence: 99%

Selective targeting of JAK/STAT signaling is potentiated by Bcl-xL blockade in IL-2–dependent adult T-cell leukemia

Zhang

Griner

Wang

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Adult T-cell leukemia (ATL) develops in individuals infected with human T-cell lymphotropic virus-1 (HTLV-1). Presently there is no curative therapy for ATL. HTLV-1-encoded protein Tax (transactivator from the X-gene region) up-regulates Bcl-xL (B-cell lymphoma-extra large) expression and activates interleukin-2 (IL-2), IL-9, and IL-15 autocrine/paracrine systems, resulting in amplified JAK/STAT signaling. Inhibition of JAK signaling reduces cytokinedependent ex vivo proliferation of peripheral blood mononuclear cells (PBMCs) from ATL patients in smoldering/chronic stages. Currently, two JAK inhibitors are approved for human use. In this study, we examined activity of multiple JAK inhibitors in ATL cell lines. The selective JAK inhibitor ruxolitinib was examined in a high-throughput matrix screen combined with >450 potential therapeutic agents, and Bcl-2/Bcl-xL inhibitor navitoclax was identified as a strong candidate for multicomponent therapy. The combination was noted to strongly activate BAX (Bcl-2-associated X protein), effect mitochondrial depolarization, and increase caspase 3/7 activities that lead to cleavage of PARP (poly ADP ribose polymerase) and Mcl-1 (myeloid cell leukemia 1). Ruxolitinib and navitoclax independently demonstrated modest antitumor efficacy, whereas the combination dramatically lowered tumor burden and prolonged survival in an ATL murine model. This combination strongly blocked ex vivo proliferation of five ATL patients' PBMCs. These studies provide support for a therapeutic trial in patients with smoldering/chronic ATL using a drug combination that inhibits JAK signaling and antiapoptotic protein Bcl-xL.adult T-cell leukemia | Janus kinase | ruxolitinib | navitoclax T -cell leukemia/lymphoma represents ∼10% of lymphoid malignancies. Genetic alterations affecting members of the Janus kinases (JAKs) and signal transducers and activators of transcription (STAT) were discovered in a variety of T-cell malignancies (1, 2). Furthermore, increases in the common γc cytokine concentrations that signal through the JAK1/3, STAT3/5 pathway have been demonstrated in angioimmunoblastic T-cell lymphoma, gamma delta T-cell lymphoma, and adult T-cell leukemia (ATL), thereby identifying effective therapeutic targets.ATL is an aggressive T-cell malignancy characterized by the clonal expansion of CD4 + CD25 + T lymphocytes that develops in a small proportion of individuals infected with human T-cell lymphotrophic virus-1 (HTLV-1) (3-5). Clinically, ATL is subclassified into four subtypes: smoldering, chronic, lymphomatous, and acute (4, 5). Presently, there is no curative therapy for ATL (4, 5). In search of effective therapies, we examined key signaling pathways that confer a proliferation and viability advantage to ATL cells. It was noted that the HTLV-1-encoded Tax (transactivator from the X-gene region) is associated with increased Bcl-xL (B-cell lymphoma-extra large) expression in ATL cells (6). Furthermore, we reported two autocrine loops (IL-2/IL-2Rα, IL-15/IL-15Rα) and one paracrine loop that in...

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Comprehensive assay of kinase catalytic activity reveals features of kinase inhibitor selectivity

Cited by 775 publications

References 35 publications

The Fibroblast Growth Factor 14·Voltage-gated Sodium Channel Complex Is a New Target of Glycogen Synthase Kinase 3 (GSK3)

The Fibroblast Growth Factor 14·Voltage-gated Sodium Channel Complex Is a New Target of Glycogen Synthase Kinase 3 (GSK3)

Targeted genetic dependency screen facilitates identification of actionable mutations in FGFR4, MAP3K9, and PAK5 in lung cancer

Selective targeting of JAK/STAT signaling is potentiated by Bcl-xL blockade in IL-2–dependent adult T-cell leukemia

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