Dynamic NF-κB and E2F interactions control the priority and timing of inflammatory signalling and cell proliferation

Ankers, John; Awais, Raheela; Jones, Nicholas A.; Boyd, James; Ryan, Sheila; Adamson, Antony; Harper, Claire V.; Bridge, Lloyd; Spiller, David G.; Jackson, Dean A.; Paszek, Pawel; Sée, Violaine; White, Mike

doi:10.7554/elife.10473

Cited by 52 publications

(56 citation statements)

References 80 publications

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“…This mechanism might also involve maintenance of the cellular NF-B concentration independently of the cell size, as we previously observed (38). Alternative mechanisms might involve a specific gating of the inflammatory response through the cell cycle system, for example, through modulation of NF-B dynamics (55). Further studies will be required to understand precisely how the observed target gene expression (and protein secretion) correlates with heterogeneous NF-B dynamics in single cells (56) and how it is influenced by other factors, for example, potential augmentation of the TLR response through paracrine or autocrine signaling (20,21).…”

Section: Discussionmentioning

confidence: 66%

Quantitative analysis of competitive cytokine signaling predicts tissue thresholds for the propagation of macrophage activation

et al. 2018

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Toll-like receptor (TLR) signaling regulates macrophage activation and effector cytokine propagation in the constrained environment of a tissue. In macrophage populations, TLR4 stimulates the dose-dependent transcription of nuclear factor κB (NF-κB) target genes. However, using single-RNA counting, we found that individual cells exhibited a wide range (three orders of magnitude) of expression of the gene encoding the proinflammatory cytokine tumor necrosis factor-α (TNF-α). The TLR4-induced transcriptional response correlated with the extent of NF-κB signaling in the cells and their size. We compared the rates of TNF-α production and uptake in macrophages and mouse embryonic fibroblasts and generated a mathematical model to explore the heterogeneity in the response of macrophages to TLR4 stimulation and the propagation of the TNF-α signal in the tissue. The model predicts that the local propagation of the TLR4-dependent TNF-α response and cellular NF-κB signaling are limited to small distances of a few cell diameters between neighboring tissue-resident macrophages. In our predictive model, TNF-α propagation was constrained by competitive uptake of TNF-α from the environment, rather than by heterogeneous production of the cytokine. We propose that the highly constrained architecture of tissues enables effective localized propagation of inflammatory cues while avoiding out-of-context responses at longer distances.

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Section: Discussionmentioning

confidence: 66%

Quantitative analysis of competitive cytokine signaling predicts tissue thresholds for the propagation of macrophage activation

et al. 2018

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“…Experiments were performed on three cell models: human neuroblastoma SK-N-AS cells transfected with RelA-dsRedXP plasmid, SK-N-AS cells stably expressing an IkBα-eGFP BAC as described previously ( 29 , 30 ), and primary mouse adult fibroblasts (MAFs) cultured from ear punches from transgenic mice expressing an RelA-DsRed XP BAC ( SI Appendix ). SK-N-AS cells were cultured in Modified Eagles Medium supplemented with 1% nonessential amino acids (Sigma-Aldrich), and MAFs were cultured in Dulbecco Modified Eagles Medium.…”

Section: Methodsmentioning

confidence: 99%

Temperature regulates NF-κB dynamics and function through timing of A20 transcription

Harper

Woodcock

Lam

et al. 2018

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

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SignificanceInflammation is often accompanied by temperature change, but little is known about the role of temperature in the inflammatory response. We show that physiologically relevant temperature changes significantly perturb NF-κB dynamics following TNFα stimulation in single cells. Using experimentation informed by mathematical modeling, we found that these changes were mediated, at least in part, through the key feedback gene TNFAIP3/A20. Curtailing A20 expression removed temperature sensitivity across the fever range (37 °C to 40 °C). Gene expression was generally unaffected between these temperatures, although a select set of NF-κB−regulated genes was up-regulated at early time points. These genes were predominantly involved in inflammation, signaling, and cell fate. The cellular response to inflammation may therefore be mechanistically and functionally regulated by temperature.

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“…Over the last decade, cutting-edge microscopy approaches have demonstrated that this two-feedback structure can support oscillations of the nuclear NF-κB in response to cytokine stimulation, including Tumor Necrosis Factor α (TNFα) [ 11 , 12 ]. These analyses exemplify how complex environmental signals may be functionally encoded in the dynamical NF-κB system response [ 13 ], which is in part facilitated by crosstalk with other cellular signaling systems [ 14 , 15 ]. For example, the treatment dose regulated the number of responding cells in the population, exhibiting digital (binary) on-off activation [ 16 – 18 ], while the timing and amplitude of the NF-κB response controls target gene expression [ 11 , 19 ].…”

Section: Introductionmentioning

confidence: 99%

Quantitative analysis reveals crosstalk mechanisms of heat shock-induced attenuation of NF-κB signaling at the single cell level

et al. 2018

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Elevated temperature induces the heat shock (HS) response, which modulates cell proliferation, apoptosis, the immune and inflammatory responses. However, specific mechanisms linking the HS response pathways to major cellular signaling systems are not fully understood. Here we used integrated computational and experimental approaches to quantitatively analyze the crosstalk mechanisms between the HS-response and a master regulator of inflammation, cell proliferation, and apoptosis the Nuclear Factor κB (NF-κB) system. We found that populations of human osteosarcoma cells, exposed to a clinically relevant 43°C HS had an attenuated NF-κB p65 response to Tumor Necrosis Factor α (TNFα) treatment. The degree of inhibition of the NF-κB response depended on the HS exposure time. Mathematical modeling of single cells indicated that individual crosstalk mechanisms differentially encode HS-mediated NF-κB responses while being consistent with the observed population-level responses. In particular “all-or-nothing” encoding mechanisms were involved in the HS-dependent regulation of the IKK activity and IκBα phosphorylation, while others involving transport were “analogue”. In order to discriminate between these mechanisms, we used live-cell imaging of nuclear translocations of the NF-κB p65 subunit. The single cell responses exhibited “all-or-nothing” encoding. While most cells did not respond to TNFα stimulation after a 60 min HS, 27% showed responses similar to those not receiving HS. We further demonstrated experimentally and theoretically that the predicted inhibition of IKK activity was consistent with the observed HS-dependent depletion of the IKKα and IKKβ subunits in whole cell lysates. However, a combination of “all-or-nothing” crosstalk mechanisms was required to completely recapitulate the single cell data. We postulate therefore that the heterogeneity of the single cell responses might be explained by the cell-intrinsic variability of HS-modulated IKK signaling. In summary, we show that high temperature modulates NF-κB responses in single cells in a complex and unintuitive manner, which needs to be considered in hyperthermia-based treatment strategies.

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Dynamic NF-κB and E2F interactions control the priority and timing of inflammatory signalling and cell proliferation

Cited by 52 publications

References 80 publications

Quantitative analysis of competitive cytokine signaling predicts tissue thresholds for the propagation of macrophage activation

Quantitative analysis of competitive cytokine signaling predicts tissue thresholds for the propagation of macrophage activation

Temperature regulates NF-κB dynamics and function through timing of A20 transcription

Quantitative analysis reveals crosstalk mechanisms of heat shock-induced attenuation of NF-κB signaling at the single cell level

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