Nucleoside Diphosphate Kinase-3 (NME3) Enhances TLR5-Induced NFκB Activation

Abstract

Bacterial flagellin is a potent activator of NFκB signaling, inflammation, and host innate immunity, and recent data indicate that flagellin represents a novel antitumor ligand acting through toll-like receptor 5 (TLR5) and the NFκB pathway to induce host immunity and aid in the clearance of tumor xenografts. To identify innate signaling components of TLR5 responsible for these antitumor effects, a loss-of-function high-throughput screen was employed utilizing carcinoma cells expressing a dynamic NFκB bioluminescent reporter stimulated by expressing flagellin. A live cell screen of a siRNA library targeting 691 known and predicted human kinases to identify novel tumor cell modulators of TLR5-induced NFκB activation uncovered several interesting positive and negative candidate regulators not previously recognized, including nucleoside diphosphate kinase 3 (NME3), characterized as an enhancer of signaling responses to flagellin. Targeted knockdown and overexpression assays confirmed the regulatory contribution of NME3 to TLR5-mediated NFκB signaling, mechanistically downstream of MyD88. Furthermore, Kaplan-Meier survival analysis showed that NME3 expression correlated highly with TLR5 expression in breast, lung, ovarian, and gastric cancers, and furthermore, high-level expression of NME3 increased overall survival for patients with breast, lung, and ovarian cancer, but the opposite in gastric cancer. Together, these data identify a previously unrecognized proinflammatory role for NME3 in signaling downstream of TLR5 that may potentiate cancer immunotherapies. Proinflammatory signaling mediated by innate immunity engagement of flagellin-activated TLR5 in tumor cells results in antitumor effects through NME3 kinase, a positive downstream regulator of flagellin-mediated NFκB signaling, enhancing survival for several human cancers. .

Therapeutic Targeting of Non-oncogene Dependencies in High-risk Neuroblastoma

“…However, its role in cancer is not fully understood. A recent report has shown that NME3 expression positively correlates with patient survival in breast or lung cancer but negatively correlates with that in gastric cancer (71). By contrast, our analysis did not reveal a dependency of high-risk neuroblastoma tumors on NME3 examined at the mRNA level ( Supplementary Table S4), but we cannot exclude the contribution of this gene product to the cancer biology.…”

“…However, owing to the lack of efficacy targets of niclosamide, it is still unable to explain how the direct binding events could relate to the decreased nucleotide biosynthesis and NME3 protein expression in neuroblastoma, as well as to the many other therapeutic effects in different disease settings. Molecularly, it has been demonstrated that NME3 can enhance NF-kB signaling downstream of Toll-like receptor 5 activation (71) and that the direct interaction of NME3 with Tip60 (also known as lysine acetyltransferase 5, or KAT5) is crucial for DNA repair (72). Further system-level studies of cellular signaling responses following niclosamide treatment are anticipated to uncover clues about the drug's efficacy targets and MoAs in different contexts.…”

Exploration of Anti-inflammatory Mechanism of Forsythiaside A and Forsythiaside B in CuSO4-Induced Inflammation in Zebrafish by Metabolomic and Proteomic Analyses

“…Besides, Nme3 was con rmed to modulate pro-in ammatory transcription by activating NF-κB [75]. This modulation activates the Tolllike receptor 5-mediated NF-κB signaling pathway in a MyD88-dependent manner [59]. In the present study, both FA and FB downregulated Nme3 expression, suggesting that FA and FB might possess a regulatory effect on cellular signaling and function.…”

“…These pathways could in uence multiple in ammation-related processes, as will be further elaborated in the discussion below on integrating the metabolomic and proteomic analyses. The changes in expression of the identi ed DEPs that are actively or passively involved in the in ammatory response, including Collagen (col2a1b, col9a2, col9a1b), Wdr3, Mrps7, and Nme3, have all been reversed both in the FA and FB groups [58][59][60][61].…”

A humanized yeast phenomic model of deoxycytidine kinase to predict genetic buffering of nucleoside analog cytotoxicity

“…NME5 overexpression was previously associated with gemcitabine-resistant cancer, and its knockdown can increase gemcitabine efficacy [182]. Thus, the anti-cancer efficacy of gemcitabine could be influenced by differential expression and activity of NDK isoforms across tissues [251], such that NME gene expression could be predictive of response to nucleoside analogs, or perhaps targeted for synergistic anti-tumor activity.…”