Mutations in UBA3 Confer Resistance to the NEDD8-Activating Enzyme Inhibitor MLN4924 in Human Leukemic Cells

Xu, Gang; Toth, Julia I.; Silva, Sara R. da; Paiva, Stacey-Lynn; Lukkarila, Julie L.; Hurren, Rose; MacLean, Neil; Sukhai, Mahadeo A.; Bhattacharjee, Rabindra N.; Goard, Carolyn A.; Gunning, Patrick T.; Dhe‐Paganon, Sirano; Petroski, Matthew D.; Schimmer, Aaron D.

doi:10.1371/journal.pone.0093530

Cited by 33 publications

(36 citation statements)

References 35 publications

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“…In tumour cell lines and xenografts, treatment-emerging resistance was observed for MLN4924. Sequencing in the resistant lines identified heterologous mutations in UBA3 (predominantly A171T) within the MgATP-binding cleft where MLN4924 also binds (Milhollen et al 2012, Toth et al 2012, Xu et al 2014. These data strongly suggest that the observed biological effects in MLN4924-treated cells are primarily due to NAE inhibition.…”

Section: Development Of Nedd8 Inhibitors-mln4924mentioning

confidence: 80%

“…Common issues for compounds that enter into clinical trials include the potential development of resistance and their use as single agents or in combination therapy. The studies in tissue culture and xenograft model systems have indeed showed that resistance to MLN4924 is a highly anticipated outcome (Milhollen et al 2012, Toth et al 2012, Xu et al 2014. These results are critical as not only demonstrate that the observed anti-tumour effects of MLN4924 depend primarily on its target, NAE, but also raise the necessity to develop new compounds that potentially will overcome resistance.…”

Section: Potential Strategies For the Use Of Nedd8 Inhibitors In Cancmentioning

confidence: 99%

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Regulation of cancer-related pathways by protein NEDDylation and strategies for the use of NEDD8 inhibitors in the clinic

Abidi

Xirodimas

2014

Endocrine-Related Cancer

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Post-translational modification of proteins with ubiquitin and ubiquitin-like molecules (UBLs) controls a vast if not every biological process in the cell. It is not surprising that deregulation in ubiquitin and UBL signalling has been implicated in the pathogenesis of many diseases and that these pathways are considered as major targets for therapeutic intervention. In this review, we summarise recent advances in our understanding of the role of the UBL neural precursor cell expressed developmentally downregulated-8 (NEDD8) in cancer-related processes and potential strategies for the use of NEDD8 inhibitors as chemotherapeutics.

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Section: Development Of Nedd8 Inhibitors-mln4924mentioning

confidence: 80%

Section: Potential Strategies For the Use Of Nedd8 Inhibitors In Cancmentioning

confidence: 99%

Regulation of cancer-related pathways by protein NEDDylation and strategies for the use of NEDD8 inhibitors in the clinic

Abidi

Xirodimas

2014

Endocrine-Related Cancer

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“…While MLN4924 has been intensively investigated in preclinical and clinical trials, the induction of drug-resistant mutations in UBA3, the molecular target of MLN4924, raises the necessity to identify other therapeutic targets against neddylation pathway [[31], [32], [33]]. In this study, we found that UBC12 was overexpressed in human lung cancer, increased with disease deterioration, and positively correlated with NEDD8 expression.…”

Section: Introductionmentioning

confidence: 81%

Validation of NEDD8-conjugating enzyme UBC12 as a new therapeutic target in lung cancer

Kang

Zhang

et al. 2019

EBioMedicine

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Background The neddylation pathway is overactivated in human cancers. Inhibition of neddylation pathway has emerged as an attractive anticancer strategy. The mechanisms underlying neddylation overactivation in cancer remain elusive. MLN4924/Pevonedistat, a first-in-class NEDD8-activating enzyme (NAE, E1) inhibitor, exerts significant anti-tumor effects, but its mutagenic resistance remains unresolved. Methods The expression of NEDD8-conjugating enzyme UBC12/UBE2M (E2) and NEDD8 were estimated by bioinformatics analysis and western blot in human lung cancer cell lines. The malignant phenotypes of lung cancer cells were evaluated both in vitro and in vivo upon UBC12 knockdown. Cell-cycle arrest was evaluated by quantitative proteomic analysis and propidium iodide stain and fluorescence - activated cell sorting (FACS). The growth of MLN4924 - resistant H1299 cells was also evaluated upon UBC12 knockdown. Findings The mRNA level of UBC12 in lung cancer tissues was much higher than that in normal lung tissues, increased with disease deterioration, and positively correlated with NEDD8 expression. Moreover, the overexpression of UBC12 significantly enhanced protein neddylation modification whereas the downregulation of UBC12 reduced neddylation modification of target proteins. Functionally, neddylation inactivation by UBC12 knockdown suppressed the malignant phenotypes of lung cancer cells both in vitro and in vivo . The quantitative proteomic analysis and cell cycle profiling showed that UBC12 knockdown disturbed cell cycle progression by triggering G 2 phase cell-cycle arrest. Further mechanistical studies revealed that UBC12 knockdown inhibited Cullin neddylation, led to the inactivation of CRL E3 ligases and induced the accumulation of tumor-suppressive CRL substrates (p21, p27 and Wee1) to induce cell cycle arrest and suppress the malignant phenotypes of lung cancer cells. Finally, UBC12 knockdown effectively inhibited the growth of MLN4924-resistant lung cancer cells. Interpretation These findings highlight a crucial role of UBC12 in fine-tuned regulation of neddylation activation status and validate UBC12 as an attractive alternative anticancer target against neddylation pathway. Fund Chinese Minister of Science and Technology grant (2016YFA0501800), National Natural Science Foundation of China (Grant Nos. 81401893, 81625018, 81820108022, 81772470, 81572340 and 81602072), Innovation Program of Shanghai Municipal Education Commission (2019-01-07-00-10-E00056), Program of Shanghai Academic/Technology Research Leader (18XD1403800), National Thirteenth Five-Year Science and Technology Major Special Project for New Drug and Development (2017ZX09304001). The funders had no role in study design, data collection, data analysis, interpretation, writing of the...

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“…In addition, other mechanisms of resistance to pevonedistat, which were not explored in this paper but should be considered particularly for cases of acquired resistance, include mutations in components of the neddylation machinery such as UBA3 (i.e. catalytic subunit of NAE) [44, 45]. …”

Section: Discussionmentioning

confidence: 99%

Targeting the protein ubiquitination machinery in melanoma by the NEDD8-activating enzyme inhibitor pevonedistat (MLN4924)

et al. 2016

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Background The neddylation pathway conjugates NEDD8 to cullin-RING ligases and controls the proteasomal degradation of specific proteins involved in essential cell processes. Pevonedistat (MLN4924) is a selective small molecule targeting the NEDD8-activating enzyme (NAE) and inhibits an early step in neddylation, resulting in DNA re-replication, cell cycle arrest and death. We investigated the anti-tumor potential of pevonedistat in preclinical models of melanoma. Methods Melanoma cell lines and patient-derived tumor xenografts (PDTX) treated with pevonedistat were assessed for viability/apoptosis and tumor growth, respectively, to identify sensitive/resistant models. Gene expression microarray and gene set enrichment analyses were performed in cell lines to determine the expression profiles and pathways of sensitivity/resistance. Pharmacodynamic changes in treated-PDTX were also characterized. Results Pevonedistat effectively inhibited cell viability (IC50 <0.3μM) and induced apoptosis in a subset of melanoma cell lines. Sensitive and resistant cell lines exhibited distinct gene expression profiles; sensitive models were enriched for genes involved in DNA repair, replication and cell cycle regulation, while immune response and cell adhesion pathways were upregulated in resistant models. Pevonedistat also reduced tumor growth in melanoma cell line xenografts and PDTX with variable responses. An accumulation of pevonedistat-NEDD8 adduct and CDT1 was observed in sensitive tumors consistent with its mechanism of action. Conclusions This study provided preclinical evidence that NAE inhibition by pevonedistat has anti-tumor activity in melanoma and supports the clinical benefits observed in recent Phase 1 trials of this drug in melanoma patients. Further investigations are warranted to develop rational combinations and determine predictive biomarkers of pevonedistat.

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Mutations in UBA3 Confer Resistance to the NEDD8-Activating Enzyme Inhibitor MLN4924 in Human Leukemic Cells

Cited by 33 publications

References 35 publications

Regulation of cancer-related pathways by protein NEDDylation and strategies for the use of NEDD8 inhibitors in the clinic

Regulation of cancer-related pathways by protein NEDDylation and strategies for the use of NEDD8 inhibitors in the clinic

Validation of NEDD8-conjugating enzyme UBC12 as a new therapeutic target in lung cancer

Targeting the protein ubiquitination machinery in melanoma by the NEDD8-activating enzyme inhibitor pevonedistat (MLN4924)

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