Treatment-Emergent Mutations in NAEβ Confer Resistance to the NEDD8-Activating Enzyme Inhibitor MLN4924

Milhollen, Michael A.; Thomas, Michael; Narayanan, Usha; Traore, Tary; Riceberg, Jessica; Amidon, Benjamin S.; Bence, Neil; Bolen, Joseph B.; Brownell, James E.; Dick, Lawrence R.; Loke, Huay-Keng; McDonald, Alice; Ma, Jingya; Manfredi, Mark; Sells, Todd B.; Sintchak, Mike; Yang, Xiaofeng; Xu, Qing; Koenig, Erik; Gavin, James M.; Smith, Peter G.

doi:10.1016/j.ccr.2012.02.009

Cited by 99 publications

(92 citation statements)

References 24 publications

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“…Our results showed that MLN4924 had significant and similar single-agent activity in both cisplatin-sensitive and -resistant models, indicating that the mechanisms that underlie resistance to cisplatin and MLN4924 likely do not overlap. This is in agreement with 2 recently published studies that identified treatment-related hetereozygous mutations in the ATP-binding pocket and NEDD8-binding cleft of NAEb as a novel mechanism of resistance to MLN4924 (12,38). Additional evidence for potentially distinct mechanisms of resistance between classical chemotherapy and MLN4924 is also provided by recent studies with MLN4924 in acute myelogenous leukemia (AML).…”

Section: Discussionsupporting

confidence: 79%

“…1B). Notably, the variation in sensitivity of individual cell lines to MLN4924 was not due to mutations in NAEb, a recently identified mechanism of resistance to MLN4924, as DNA sequencing of all cell lines used in this study failed to detect any mutations (12).…”

Section: Mln4924 Diminishes Cell Viability and Impairs The Clonogenicmentioning

confidence: 93%

“…DNA was eluted with 100 mL nuclease-free water and samples were checked for concentration and quality using a NanoDrop spectrophotometer. PCR amplifications were conducted using optimized cycling conditions per gene-exon following the previously described protocol (12). All samples were sequenced with forward and reverse primers to obtain the…”

Section: Naeb Dna Sequencingmentioning

confidence: 99%

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Disrupting Protein NEDDylation with MLN4924 Is a Novel Strategy to Target Cisplatin Resistance in Ovarian Cancer

Nawrocki

Kelly

Smith

et al. 2013

Clinical Cancer Research

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Purpose: Ovarian cancer has the highest mortality rate of all female reproductive malignancies. Drug resistance is a major cause of treatment failure and novel therapeutic strategies are urgently needed. MLN4924 is a NEDDylation inhibitor currently under investigation in multiple phase I studies. We investigated its anticancer activity in cisplatin-sensitive and -resistant ovarian cancer models.Experimental Design: Cellular sensitivity to MLN4924/cisplatin was determined by measuring viability, clonogenic survival, and apoptosis. The effects of drug treatment on global protein expression, DNA damage, and reactive oxygen species generation were determined. RNA interference established natural born killer/bcl-2-interacting killer (NBK/BIK) as a regulator of therapeutic sensitivity. The in vivo effects of MLN4924/cisplatin on tumor burden and key pharmacodynamics were assessed in cisplatin-sensitive and -resistant xenograft models.Results: MLN4924 possessed significant activity against both cisplatin-sensitive and -resistant ovarian cancer cells and provoked the stabilization of key NEDD8 substrates and regulators of cellular redox status. Notably, MLN4924 significantly augmented the activity of cisplatin against cisplatin-resistant cells, suggesting that aberrant NEDDylation may contribute to drug resistance. MLN4924 and cisplatin cooperated to induce DNA damage, oxidative stress, and increased expression of the BH3-only protein NBK/BIK. Targeted NBK/BIK knockdown diminished the proapoptotic effects of the MLN4924/cisplatin combination. Administration of MLN4924 to mice bearing ovarian tumor xenografts significantly increased the efficacy of cisplatin against both cisplatin-sensitive and -resistant tumors.Conclusions: Our collective data provide a rationale for the clinical investigation of NEDD8-activating enzyme (NAE) inhibition as a novel strategy to augment cisplatin efficacy in patients with ovarian cancer and other malignancies.

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Section: Discussionsupporting

confidence: 79%

Section: Mln4924 Diminishes Cell Viability and Impairs The Clonogenicmentioning

confidence: 93%

Section: Naeb Dna Sequencingmentioning

confidence: 99%

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Disrupting Protein NEDDylation with MLN4924 Is a Novel Strategy to Target Cisplatin Resistance in Ovarian Cancer

Nawrocki

Kelly

Smith

et al. 2013

Clinical Cancer Research

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“…Nedd8 conditional KO also did not yield a productive model, since Nedd8 is required for cellular homeostasis. Therefore, in order to validate the results obtained with RNAi technology and establish the in vivo physiological relevance of neddylation during IL-1␤ production, we utilized an inhibitor of neddylation called MLN4924, which blocks the activity of Nedd8-activating enzyme (47)(48)(49). MLN4924 is currently being developed as an anticancer drug during tumorogenesis, since neddylation of the tumor suppressor protein p53 results in loss of its activity.…”

Section: Resultsmentioning

confidence: 99%

“…This was due to diminished processing of pro-caspase-1 to yield functional subunits (i.e., the p10 subunit). The knockdown studies were further validated by using MLN4924, a neddylation inhibitor that blocks the activity of Nedd8-activating enzyme (NAE) (47)(48)(49). Further studies revealed colocalization and coimmunoprecipitation of endogenous Nedd8 with caspase-1 in inflammasome-activated macrophages.…”

mentioning

confidence: 99%

Nedd8 Regulates Inflammasome-Dependent Caspase-1 Activation

Segovia

Tsai

Chang

et al. 2015

Molecular and Cellular Biology

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dCaspase-1 is activated by the inflammasome complex to process cytokines like interleukin-1␤ (IL-1␤). Pro-caspase-1 consists of three domains, CARD, p20, and p10. Association of pro-caspase-1 with the inflammasome results in initiation of its autocatalytic activity, culminating in self-cleavage that generates catalytically active subunits (p10 and p20). In the current study, we show that Nedd8 is required for efficient self-cleavage of pro-caspase-1 to generate its catalytically active subunits. Nedd8 silencing or treating cells with the neddylation inhibitor MLN4924 led to diminished caspase-1 processing and reduced IL-1␤ maturation following inflammasome activation. Coimmunoprecipitation and mass spectrometric analysis of 293 cells overexpressing procaspase-1 (and CARD) and Nedd8 suggested possible neddylation of caspase-1 CARD. Following inflammasome activation in primary macrophages, we observed colocalization of endogenous Nedd8 with caspase-1. Similarly, interaction of endogenous Nedd8 with caspase-1 CARD was detected in inflammasome-activated macrophages. Furthermore, enhanced autocatalytic activity of pro-caspase-1 was observed following Nedd8 overexpression in 293 cells, and such activity in inflammasome-activated macrophages was drastically diminished upon treatment of cells with MLN4924. Thus, our studies demonstrate a role of Nedd8 in regulating caspase-1 activation following inflammasome activation, presumably via augmenting autoprocessing/cleavage of pro-caspase-1 into its corresponding catalytically active subunits.

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Neddylation inhibition upregulates PD‐L1 expression and enhances the efficacy of immune checkpoint blockade in glioblastoma

Zhou

Zhao

Yang

et al. 2019

Intl Journal of Cancer

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Pevonedistat (MLN4924), a specific NEDD8‐activating enzyme inhibitor, has been considered as a promising treatment for glioblastoma, which is currently in Phase I/II clinical trials. On the other hand, inhibition of neddylation pathway substantially upregulates the expression of T cell negative regulator programmed death‐ligand 1 (PD‐L1), which might account for the potential resistance via evasion of immune surveillance checkpoints. Whether administration of anti‐PD‐L1 enhances the efficacy of pevonedistat through a cytotoxic T cell‐dependent mechanism in glioblastoma needs to be investigated. Here, we report that depletion of neddylation pathway key enzymes markedly elevates PD‐L1 expression in glioblastoma cancer cells. Consistently, neddylation inhibitor pevonedistat significantly enhances PD‐L1 expression in both glioblastoma cancer cell lines and animal models. Mechanistically, pevonedistat increases PD‐L1 mRNA levels mainly through inhibiting Cullin1‐F‐box and WD repeat domain‐containing 7 E3 ligase activity and accumulating c‐MYC proteins, a direct transcriptional activator of PD‐L1 gene expression. In addition, inhibition of Cullin3 activity by pevonedistat also blocks PD‐L1 protein degradation. Importantly, pevonedistat attenuates T cell killing through PD‐L1 induction, and blockade of PD‐L1 restores the sensitivity of pevonedistat‐treated glioblastoma cancer cells to T cell killing. The combination of pevonedistat and anti‐PD‐L1 therapy compared to each agent alone significantly increased the therapeutic efficacy in vivo. Our study demonstrates inhibition of neddylation pathway suppresses cancer‐associated immunity and provides solid evidence to support the combination of pevonedistat and PD‐L1/programmed cell death protein 1 immune checkpoint blockade as a potential therapeutic strategy to treat glioblastoma.

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Treatment-Emergent Mutations in NAEβ Confer Resistance to the NEDD8-Activating Enzyme Inhibitor MLN4924

Cited by 99 publications

References 24 publications

Disrupting Protein NEDDylation with MLN4924 Is a Novel Strategy to Target Cisplatin Resistance in Ovarian Cancer

Disrupting Protein NEDDylation with MLN4924 Is a Novel Strategy to Target Cisplatin Resistance in Ovarian Cancer

Nedd8 Regulates Inflammasome-Dependent Caspase-1 Activation

Neddylation inhibition upregulates PD‐L1 expression and enhances the efficacy of immune checkpoint blockade in glioblastoma

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