Escalating Threat Levels of Bacterial Infection Can Be Discriminated by Distinct MAPK and NF-κB Signaling Dynamics in Single Host Cells

Lane, Keara; Andres-Terrè, Marta; Kudo, Tetsuichi; Monack, Denise M.; Covert, Markus W.

doi:10.1016/j.cels.2019.02.008

Cited by 26 publications

(25 citation statements)

References 72 publications

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“…Comparing invasion by WT bacteria to invasion by a mutant that is defective for intracellular survival due to disruption of the global regulator of virulence AlgR, we found that the host NF-κB response is stronger in WT-invaded compared to ΔalgR -invaded cells. A recent study showed that high and sustained NF-κB signaling and maximal JNK signaling only occurred in response to live bacteria that were sensed by the host as a threat, as opposed to heat-killed bacteria [ 63 ], suggesting that the relatively muted NF-κB signaling on the part of the host in response to ΔalgR invasion may be because it is not sensing the same threat level as it does from WT bacteria. Paradoxically, while the NF-κB response plays a role in clearing of extracellular bacteria [ 17 ], it appears to be required for intracellular bacterial survival, as its inhibition with IKK inhibitors leads to decreased survival of intracellular WT bacteria.…”

Section: Discussionmentioning

confidence: 99%

Dual transcriptional analysis reveals adaptation of host and pathogen to intracellular survival of Pseudomonas aeruginosa associated with urinary tract infection

2021

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Long-term survival of bacterial pathogens during persistent bacterial infections can be associated with antibiotic treatment failure and poses a serious public health problem. Infections caused by the Gram-negative pathogen Pseudomonas aeruginosa, which can cause both acute and chronic infections, are particularly challenging due to its high intrinsic resistance to antibiotics. The ineffectiveness of antibiotics is exacerbated when bacteria reside intracellularly within host cells where they can adopt a drug tolerant state. While the early steps of adherence and entry of P. aeruginosa into mammalian cells have been described, the subsequent fate of internalized bacteria, as well as host and bacterial molecular pathways facilitating bacterial long-term survival, are not well defined. In particular, long-term survival within bladder epithelial cells has not been demonstrated and this may have important implications for the understanding and treatment of UTIs caused by P. aeruginosa. Here, we demonstrate and characterize the intracellular survival of wild type (WT) P. aeruginosa inside bladder epithelial cells and a mutant with a disruption in the bacterial two-component regulator AlgR that is unable to survive intracellularly. Using simultaneous dual RNA-seq transcriptional profiling, we define the transcriptional response of intracellular bacteria and their corresponding invaded host cells. The bacterial transcriptional response demonstrates that WT bacteria rapidly adapt to the stress encountered in the intracellular environment in contrast to ΔalgR bacteria. Analysis of the host transcriptional response to invasion suggests that the NF-κB signaling pathway, previously shown to be required for extracellular bacterial clearance, is paradoxically also required for intracellular bacterial survival. Lastly, we demonstrate that intracellular survival is important for pathogenesis of P. aeruginosa in vivo using a model of murine urinary tract infection. We propose that the unappreciated ability of P. aeruginosa to survive intracellularly may play an important role in contributing to the chronicity and recurrence of P. aeruginosa in urinary tract infections.

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Section: Discussionmentioning

confidence: 99%

Dual transcriptional analysis reveals adaptation of host and pathogen to intracellular survival of Pseudomonas aeruginosa associated with urinary tract infection

2021

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“…Innate immune cells use pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs), to detect molecules that are indicative of an infection. A cell line expressing both an NF-κB and a c-Jun N-terminal kinase (JNK) reporter was challenged with increasing levels of immune stimulation, from only LPS to infection with Salmonella typhimirium , allowing for measurements of the cell's signaling response in each case (35). This work showed how an individual cell uses TLR signaling to discriminate similar signals in a variety of different contexts.…”

Section: Dynamic Decoding Of Cellular Informationmentioning

confidence: 99%

“…For example, in a population of cells exposed to S . typhimirium , uninfected cells typically activated only NF-κB, while those cells that were infected with bacteria typically activated both JNK and NF-κB (35).…”

Section: Dynamic Decoding Of Cellular Informationmentioning

confidence: 99%

Techniques for Studying Decoding of Single Cell Dynamics

2019

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Cells must be able to interpret signals they encounter and reliably generate an appropriate response. It has long been known that the dynamics of transcription factor and kinase activation can play a crucial role in selecting an individual cell's response. The study of cellular dynamics has expanded dramatically in the last few years, with dynamics being discovered in novel pathways, new insights being revealed about the importance of dynamics, and technological improvements increasing the throughput and capabilities of single cell measurements. In this review, we highlight the important developments in this field, with a focus on the methods used to make new discoveries. We also include a discussion on improvements in methods for engineering and measuring single cell dynamics and responses. Finally, we will briefly highlight some of the many challenges and avenues of research that are still open.

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“…To explore how generally applicable FRA is, we examined responses to cytokines IFN-γ, IL-10, and TNF-α 4,27 . As IFN-γ and IL-10 are implicated in macrophage phenotypic diversity 32 , we used the human monocyte cell line U937, differentiated into macrophage-like cells, and immunostaining to measure responses via nuclear levels of the key signaling effectors, phosphorylated STAT1 for IFN-γ, and phosphorylated STAT3 for IL-10 at 30 minutes after stimulation ( Fig.…”

Section: Logarithmic Dose Responses Are a General Property Of Cytokinmentioning

confidence: 99%

“…For instance, low doses of TNF-α may induce one peak of nuclear factor-κB (NF-κB) signaling activity, whereas higher doses may induce additional peaks 7,24 . Besides, the dose may control the onset, shut off, amplitude, or, in principle, any other characteristics of the responses [25][26][27][28] . Overall, conventional dose-response curves do not capture the inherent complexity of single-cell high-throughput data, and an alternative approach is required.…”

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confidence: 99%

Fractional response analysis reveals logarithmic cytokine responses in cellular populations

Nienałtowski

Rigby

Walczak

et al. 2020

Preprint

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Although we can now measure single-cell signaling responses with multivariate, high-throughput techniques our ability to interpret such measurements is still limited. Even interpretation of dose-response based on single-cell data is not straightforward: signaling responses can differ significantly between cells, encompass multiple signaling effectors, and have dynamic character. Here, we use probabilistic modeling and information-theory to introduce fractional response analysis (FRA), which quantifies changes in fractions of cells with given response levels. FRA can be universally performed for heterogeneous, multivariate, and dynamic measurements and, as we demonstrate, uncovers otherwise hidden patterns in single-cell data. In particular, we show that fractional responses to type I interferon in human peripheral blood mononuclear cells are very similar across different cell types, despite significant differences in mean or median responses and degrees of cell-to-cell heterogeneity. Further, we demonstrate that fractional responses to cytokines scale linearly with the log of the cytokine dose, which uncovers that cellular populations are sensitive to fold-changes in the dose, as opposed to additive changes.

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Escalating Threat Levels of Bacterial Infection Can Be Discriminated by Distinct MAPK and NF-κB Signaling Dynamics in Single Host Cells

Abstract: Highlights d Live-cell imaging reveals heterogeneity in host signaling during infection d NF-kB and JNK dynamics vary with bacterial location, pathogenicity, and replication d Activation of JNK is a signature of an escalation in bacterial threat level

Cited by 26 publications

References 72 publications

Dual transcriptional analysis reveals adaptation of host and pathogen to intracellular survival of Pseudomonas aeruginosa associated with urinary tract infection

Dual transcriptional analysis reveals adaptation of host and pathogen to intracellular survival of Pseudomonas aeruginosa associated with urinary tract infection

Techniques for Studying Decoding of Single Cell Dynamics

Fractional response analysis reveals logarithmic cytokine responses in cellular populations

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