Characteristic ERK1/2 signaling dynamics distinguishes necroptosis from apoptosis

Sipieter, François; Cappe, Benjamin; Leray, Aymeric; Schutter, Elke De; Bridelance, Jolien; Hulpiau, Paco; Camp, Guy Van; Declercq, Wim; Héliot, Laurent; Vincent, Pierre; Vandenabeele, Peter; Riquet, Franck B.

doi:10.1016/j.isci.2021.103074

Cited by 11 publications

(12 citation statements)

References 65 publications

(84 reference statements)

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“…We ask to what extent signaling and morphology changes contribute to heterogeneous and dynamic chemotactic movement. We found that cells also exhibit fluctuations in ERK and Akt signaling, similar to what were reported in previous studies, driving cell behaviors like cell division, death, and migration (12,13,(16)(17)(18)(19)(20). Single-cell signaling responses varied from strong fluctuations of both ERK and Akt to undetectable fluctuations (Fig.…”

Section: A-ii)supporting

confidence: 87%

Cell-to-cell variability of dynamic CXCL12-CXCR4 signaling and morphological processes in chemotaxis

Srivastava

Kinnunen

et al. 2022

Preprint

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Chemotaxis drives critical processes in cancer metastasis. While commonly studied at the population scale, metastasis arises from small numbers of cells that successfully disseminate, underscoring the need to analyze chemotaxis at single-cell resolution. Here we focus on chemotaxis driven by the CXCL12-CXCR4 pathway, a signaling network that promotes metastasis in more than 20 different human cancers. CXCL12-CXCR4 activates ERK and Akt, kinases known to promote chemotaxis, but how cells couple signaling to chemotaxis remain poorly defined. To address this challenge, we implemented single-cell analysis of MDA-MB-231 breast cancer cells migrating in a chemotaxis device towards chemokine CXCL12. We integrated live, single-cell imaging with advanced computational analysis methods to discover processes defining subsets of cells that move efficiently toward a CXCL12 gradient. We identified dynamic oscillations in ERK and Akt signaling and associated morphological transitions as key determinants of successful chemotaxis. Cells with effective chemotaxis toward CXCL12 exhibit faster and more persistent movement than non-migrating cells, but both cell populations show similar random motion. Migrating cells exhibit higher amplitude fluctuations in ERK and Akt signaling and greater frequencies of generating lateral cell membrane protrusions. Interestingly, computational analysis reveals less correlated network coupling of signaling and morphological changes in migrating cells. These data reveal processing events that enable cells to convert a signaling input to chemotaxis and highlight how cells in a uniform environment produce heterogeneous responses.

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Section: A-ii)supporting

confidence: 87%

Cell-to-cell variability of dynamic CXCL12-CXCR4 signaling and morphological processes in chemotaxis

Srivastava

Kinnunen

et al. 2022

Preprint

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“…The Erk1/2 pathway was reported to play a critical role in determining cellular responses, such as proliferation and differentiation, and phosphorylated Erk1/2 was involved in tumor progression. − As a downstream regulator of the Erk1/2 pathway, COX-2 plays an important role in the genesis and development of cancer. − As downstream genes regulated by RARβ, the protein expressions of the total Erk1/2 (t-Erk1/2), phosphorylated Erk1/2 (p-Erk1/2), and COX-2 in H4IIE cells were measured. In the absence of RA (an RARβ agonist) and U0126 (an Erk1/2 antagonist), there was no significant modification in the total Erk1/2 protein; however, the expression of p-Erk1/2 was changed (Figure A).…”

Section: Resultsmentioning

confidence: 99%

Carcinogenic Risk of 2,6-Di-tert-Butylphenol and Its Quinone Metabolite 2,6-DTBQ Through Their Interruption of RARβ: In Vivo, In Vitro, and In Silico Investigations

Cui

Zhan

et al. 2021

Environ. Sci. Technol.

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Thousands of contaminants are used worldwide and eventually released into the environment, presenting a challenge of health risk assessment. The identification of key toxic pathways and characterization of interactions with target biomacromolecules are essential for health risk assessments. The adverse outcome pathway (AOP) incorporates toxic mechanisms into health risk assessment by emphasizing the relationship among molecular initiating events (MIEs), key events (KEs), and adverse outcome (AO). Herein, we attempted the use of AOP to decipher the toxic effects of 2,6-di-tert-butylphenol (2,6-DTBP) and its para-quinone metabolite 2,6-di-tert-butyl-1,4-benzoquinone (2,6-DTBQ) based on integrated transcriptomics, molecular modeling, and cell-based assays. Through transcriptomics and quantitative real-time PCR validation, we identified retinoic acid receptor β (RARβ) as the key target biomacromolecule. The epigenetic analysis and molecular modeling revealed RARβ interference as one MIE, including DNA methylation and conformational changes. In vitro assays extended subsequent KEs, including altered protein expression of p-Erk1/2 and COX-2, and promoted cancer cell H4IIE proliferation and metastasis. These toxic effects altogether led to carcinogenic risk as the AO of 2,6-DTBP and 2,6-DTBQ, in line with chemical carcinogenesis identified from transcriptome profiling. Overall, our simplified AOP network of 2,6-DTBP and 2,6-DTBQ facilitates relevant health risk assessment.

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“…This difference in inhibition kinetics could reflect differences in conformational flexibility and/or steric hindrance that reduce accessibility of the sensor backbone to phosphatases. 64,65 We also found that the JF549SNAP/JF646Halo combination offered a 2-fold higher contrast than the JF549Halo/ JF646SNAP combination, suggesting that in at least some systems the labeling efficiency of the Halo domains is higher than that of the SNAP domains.…”

Section: ■ Results and Discussionmentioning

confidence: 65%

“…In this paper, we evaluated this combination by replacing the FP moieties in existing sensors with SNAP and Halo domains (Figure 1) and examining the performance of the modified sensors after live-cell staining with a range of organic dyes. 63 In particular, we found that we could readily produce chemigenetic variants of AKAR3EV, 64 a sensor for protein kinase A (PKA), EKAR4.0, 65 an extracellular signal-regulated kinase (ERK) activity reporter, and Yellow Cameleon 3.6 (YC), a sensor for Ca 2+ . 66 The resulting constructs can deliver similar signal-to-noise ratios compared to their CFP/YFPbased ancestors, yet permit convenient spectral tunability that readily allows the creation of color-shifted variants.…”

mentioning

confidence: 99%

Spectrally Tunable Förster Resonance Energy Transfer-Based Biosensors Using Organic Dye Grafting

et al. 2022

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Biosensors based on Förster resonance energy transfer (FRET) have revolutionized cellular biology by allowing the direct measurement of biochemical processes in situ. Many genetically encoded sensors make use of fluorescent proteins that are limited in spectral versatility and that allow few ways to change the spectral properties once the construct has been created. In this work, we developed genetically encoded FRET biosensors based on the chemigenetic SNAP and HaloTag domains combined with matching organic fluorophores. We found that the resulting constructs can display comparable responses, kinetics, and reversibility compared to their fluorescent protein-based ancestors, but with the added advantage of spectral versatility, including the availability of red-shifted dye pairs. However, we also find that the introduction of these tags can alter the sensor readout, showing that careful validation is required before applying such constructs in practice. Overall, our approach delivers an innovative methodology that can readily expand the spectral variety and versatility of FRET-based biosensors.

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Characteristic ERK1/2 signaling dynamics distinguishes necroptosis from apoptosis

Cited by 11 publications

References 65 publications

Cell-to-cell variability of dynamic CXCL12-CXCR4 signaling and morphological processes in chemotaxis

Cell-to-cell variability of dynamic CXCL12-CXCR4 signaling and morphological processes in chemotaxis

Carcinogenic Risk of 2,6-Di-tert-Butylphenol and Its Quinone Metabolite 2,6-DTBQ Through Their Interruption of RARβ: In Vivo, In Vitro, and In Silico Investigations

Spectrally Tunable Förster Resonance Energy Transfer-Based Biosensors Using Organic Dye Grafting

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