Destabilizing NEK2 overcomes resistance to proteasome inhibition in multiple myeloma

Franqui-Machin, Reinaldo; Hao, Mu; Bai, Hua; Gu, Zhimin; Zhan, Xin; Habelhah, Hasem; Jethava, Yogesh; Qiu, Lugui; Frech, Ivana; Tricot, Guido; Zhan, Fenghuang

doi:10.1172/jci98765

Cited by 68 publications

(72 citation statements)

References 71 publications

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“…These data suggested that NEK2 might elevate Beclin‐1 through blocking proteasomal degradation. We have reported that USP7 binds and stabilizes NEK2 protein (Franqui‐Machin et al , ). NEK2 interacts with both USP7 and Beclin‐1 in MM cells, and we thus hypothesized that NEK2 stabilizes Beclin‐1 via interacting with USP7.…”

Section: Resultsmentioning

confidence: 99%

“…USP7 binds to NEK2 and prevents NEK2 ubiquitination resulting in NEK2 stabilization. Increased NEK2 activates the canonical NF‐κB signaling pathway through the PP1α/AKT axis, which is the downstream targets of USP7‐NEK2 axis (Franqui‐Machin et al , ). In this study, we showed knockdown of Beclin‐1 prevents NEK2‐mediated BTZ resistance in MM both in vitro and in vivo .…”

Section: Discussionmentioning

confidence: 99%

“…Subsequent mechanistic studies showed that NEK2 induces drug resistance through up‐regulation of efflux drug pumps in MM cells. NEK2 was also found to bind to USP7, a deubiquitinase that contributes to malignancy and BTZ resistance, and was stabilized by USP7 (Franqui‐Machin et al , ). Destabilization of NEK2 by USP7 inhibitor was able to overcome resistance to BTZ in MM cells.…”

Section: Introductionmentioning

confidence: 99%

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NEK2 induces autophagy‐mediated bortezomib resistance by stabilizing Beclin‐1 in multiple myeloma

Xia

Meng³

et al. 2020

Molecular Oncology

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NEK2 is associated with drug resistance in multiple cancers. Our previous studies indicated that high NEK2 confers inferior survival in multiple myeloma (MM); thus, a better understanding of the mechanisms by which NEK2 induces drug resistance in MM is required. In this study, we discovered that NEK2 enhances MM cell autophagy, and a combination of autophagy inhibitor chloroquine (CQ) and chemotherapeutic bortezomib (BTZ) significantly prevents NEK2‐induced drug resistance in MM cells. Interestingly, NEK2 was found to bind and stabilize Beclin‐1 protein but did not affect its mRNA expression and phosphorylation. Moreover, autophagy enhanced by NEK2 was significantly prevented by knockdown of Beclin‐1 in MM cells, suggesting that Beclin‐1 mediates NEK2‐induced autophagy. Further studies demonstrated that Beclin‐1 ubiquitination is decreased through NEK2 interaction with USP7. Importantly, knockdown of Beclin‐1 sensitized NEK2‐overexpressing MM cells to BTZ in vitro and in vivo. In conclusion, we identify a novel mechanism whereby autophagy is activated by the complex of NEK2/USP7/Beclin‐1 in MM cells. Targeting the autophagy signaling pathway may provide a promising therapeutic strategy to overcome NEK2‐induced drug resistance in MM.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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NEK2 induces autophagy‐mediated bortezomib resistance by stabilizing Beclin‐1 in multiple myeloma

Xia

Meng³

et al. 2020

Molecular Oncology

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“…Among the DUBs we tested, USP7 has the greatest number of identified substrates, most of which are targeted to the proteasome 61 . Among the substrates that we identified, a few such as MARCH7, SVIL, NEK2A and TRIP12 are known targets of USP7 29, 44–46 whereas two others, PIM3 and WASL, are likely USP7 targets as USP7 is known to regulate related proteins (PIM2 47 and WASH 48 respectively) in human cells.…”

Section: Discussionmentioning

confidence: 99%

“…Our approach began with identifying proteins protected from proteasomal degradation by cysteine-protease DUBs. Several of these proteins are known to be regulated by specific DUBs, including MARCH7, BIRC3, STAM, NFX1,TRIP12, SVIL, and NEK2A 29–32, 44–46 . Furthermore, PIM3 and WASL have not been connected previously to DUBs, but are similar in sequence to well-established DUB substrates (PIM2 47 and WASH 48 respectively).…”

Section: Discussionmentioning

confidence: 99%

Proteomics of broad deubiquitylase inhibition unmasks redundant enzyme function to reveal substrates

Rossio

Paulo

Chick

et al. 2019

Preprint

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1Deubiquitylating enzymes (DUBs) counteract ubiquitylation to control the stability or activity of 2 their substrates. Identifying DUB substrates is challenging and genetic approaches can be 3 thwarted by redundant action of DUBs. Here, we circumvented redundancy by broadly inhibiting 4 DUBs in Xenopus egg extract and used quantitative mass spectrometry to identify over thirty 5 proteins that undergo proteasomal degradation, the majority of which have not been reported as 6 DUB substrates. These results were confirmed with recombinant human proteins, demonstrating 7 the conservation of their DUB-dependent stability. We used these substrates to profile the ability 8 of a panel of DUBs to rescue degradation. This approach revealed that USP7, uniquely among 9 the 14 DUBs tested, has a broad ability to rescue degradation. USP21, which is used widely to 10 nonspecifically deubiquitylate proteins in vitro, was unable to rescue degradation, highlighting 11 the importance of profiling enzyme activity in a physiological system. Together, we identify new 12 DUB substrates and present a system to characterize physiological DUB specificity, overcoming 13 the challenges posed by DUB redundancy. 14 2 Here, we first took an unbiased quantitative proteomic approach and discovered new 53 substrates protected from proteasomal degradation by DUBs, using the Xenopus egg extract 54 model system. By broadly inhibiting cysteine-protease DUBs, we circumvented the possible 55 effects of their redundancy on proteome stability. Because transcription and translation are not 56 active in this system, protein instability caused by DUB inhibition is not a consequence of 57 protein quality control ubiquitination activity. We recapitulated protein destabilization with 58 expression of recombinant human orthologs in extract, demonstrating that DUB-dependent 59 4 protein stability is evolutionarily conserved. Next, we took advantage of these newly identified 60 DUB substrates and tested the ability of a set of DUBs to rescue their degradation, uncovering a 61 broad ability of USP7 to rescue protein instability in extract. However, specific inhibition of 62 USP7 with a small molecule inhibitor was not sufficient to promote degradation of most of the 63 substrates we identified, suggesting that USP7 functions redundantly with other DUBs in this 64 system. Our work highlights the impact of DUB redundancy on proteome stability and reveals 65 the specificity and activity of DUBs whose function would otherwise be masked by redundancy. 66 67Results 68 UbVS treatment induces rapid depletion of available ubiquitin and labels a broad set of 69 cysteine-protease DUBs in Xenopus extract 70Our laboratory previously showed that the broad cysteine-protease DUB inhibitor UbVS 71 induces depletion of available ubiquitin in Xenopus extract 22 . In particular, UbVS-treatment 72 inhibited proteasome degradation of cyclin B, which could be efficiently rescued by addition of 73 50 µM exogenous ubiquitin 22 . We confirmed that 10 µM UbVS was sufficient to rapidly deplet...

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Deubiquitylating Enzymes in Cancer and Immunity

Ren,

Yu,

Liu

et al. 2023

Advanced Science

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Deubiquitylating enzymes (DUBs) maintain relative homeostasis of the cellular ubiquitome by removing the post‐translational modification ubiquitin moiety from substrates. Numerous DUBs have been demonstrated specificity for cleaving a certain type of ubiquitin linkage or positions within ubiquitin chains. Moreover, several DUBs perform functions through specific protein–protein interactions in a catalytically independent manner, which further expands the versatility and complexity of DUBs’ functions. Dysregulation of DUBs disrupts the dynamic equilibrium of ubiquitome and causes various diseases, especially cancer and immune disorders. This review summarizes the Janus‐faced roles of DUBs in cancer including proteasomal degradation, DNA repair, apoptosis, and tumor metastasis, as well as in immunity involving innate immune receptor signaling and inflammatory and autoimmune disorders. The prospects and challenges for the clinical development of DUB inhibitors are further discussed. The review provides a comprehensive understanding of the multi‐faced roles of DUBs in cancer and immunity.

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Destabilizing NEK2 overcomes resistance to proteasome inhibition in multiple myeloma

Cited by 68 publications

References 71 publications

NEK2 induces autophagy‐mediated bortezomib resistance by stabilizing Beclin‐1 in multiple myeloma

NEK2 induces autophagy‐mediated bortezomib resistance by stabilizing Beclin‐1 in multiple myeloma

Proteomics of broad deubiquitylase inhibition unmasks redundant enzyme function to reveal substrates

Deubiquitylating Enzymes in Cancer and Immunity

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