A20 prevents inflammasome-dependent arthritis by inhibiting macrophage necroptosis through its ZnF7 ubiquitin-binding domain

Polykratis, Apostolos; Martens, Arne; Eren, Remzi Onur; Shirasaki, Yoshitaka; Yamagishi, Mai; Yamaguchi, Yoshifumi; Uemura, Shunsuke; Miura, Masayuki; Holzmann, Bernhard; Kollias, George; Armaka, Marietta; Loo, Geert; Pasparakis, Manolis

doi:10.1038/s41556-019-0324-3

Cited by 124 publications

(129 citation statements)

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“…In line with this model, Gsdmd deficiency severely abrogates IL‐1β secretion upon canonical inflammasome activation (Kayagaki et al , ; Shi et al , ); and single‐cell analysis of macrophages revealed that IL‐1β release coincides with the uptake of membrane‐impermeable nucleic acid dyes (e.g. SYTOX, propidium iodide), a widely used assay to measure the loss of plasma membrane integrity (Liu et al , ; Polykratis et al , ). By contrast, a number of studies reported that mature IL‐1β can be secreted in the absence of intracellular lactate dehydrogenase release, a commonly used assay to quantify cell lysis in a bulk cell population (Kang et al , ; Chen et al , ; Gaidt et al , ; Wolf et al , ; Zanoni et al , ).…”

Section: Introductionmentioning

confidence: 90%

Beyond inflammasomes: emerging function of gasdermins during apoptosis and NETosis

2019

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Programmed cell death is a key mechanism involved in several biological processes ranging from development and homeostasis to immunity, where it promotes the removal of stressed, damaged, malignant or infected cells. Abnormalities in the pathways leading to initiation of cell death or removal of dead cells are consequently associated with a range of human diseases including infections, autoinflammatory disease, neurodegenerative disease and cancer. Apoptosis, pyroptosis and NETosis are three well-studied modes of cell death that were traditionally believed to be independent of one another, but emerging evidence indicates that there is extensive cross-talk between them, and that all three pathways can converge onto the activation of the same cell death effector-the pore-forming protein Gasdermin D (GSDMD). In this review, we highlight recent advances in gasdermin research, with a particular focus on the role of gasdermins in pyroptosis, NETosis and apoptosis, as well as cell type-specific consequences of gasdermin activation. In addition, we discuss controversies surrounding a related gasdermin family protein, Gasdermin E (GSDME), in mediating pyroptosis and secondary necrosis following apoptosis, chemotherapy and inflammasome activation.Pore-forming activity and structural autoinhibition of the gasdermin family. Nature 535: 111 -116

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Section: Introductionmentioning

confidence: 90%

Beyond inflammasomes: emerging function of gasdermins during apoptosis and NETosis

2019

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“…The tnfaip3 zebrafish orthologue encoding a predicted 762 amino acid polypeptide comprising an N-terminal ovarian tumour (OTU) domain (identity score of 94/100), including the putative catalytic cysteine at position 103 in A20’s OTU domain 28 , and seven zinc finger motifs in the C-terminus (identity scores of 79/100) (Figure S2). The zebrafish A20 C-terminal domain also contained zinc finger 4 (ZnF4) and ZnF7 regions, which showed multiple sequence alignment scores of 91/100 and 100/100 respectively, and are thus highly homologous to functional ZnF4 and ZnF7 domains identified in mammalian studies 17,21 . In mammalian cells A20 expression is controlled at the level of transcription by the NF-κB pathway in response to microbial stimulation of TLR 19 , a pathway that represents a key component of innate immunity conserved in zebrafish 29 .…”

Section: Resultsmentioning

confidence: 93%

“…A20 loss of function mutations result in macrophage hyper responsiveness to microbial stimulation and spontaneous inflammasome activation 9,13,17 . Tissue macrophages play a key role in sensing pathogenic microbiota in zebrafish 33,35 .…”

Section: Resultsmentioning

confidence: 99%

“…The A20 zinc finger 4 (ZnF4) exhibits E3 ligase activity, adding K48-linked ubiquitin chains to RIPK1, triggering RIPK1 proteolysis 12,16 . In addition, the C-terminal ZnF7 domain of A20 binds linear ubiquitin to non-catalytically suppress NF-κB activity 17 . A20 is regulated at the level of gene transcription 6 , whereby NF-κB activation induces TNFAIP3 expression 18,19 forming a negative feedback loop, and the inhibitory activities of A20 enzymatic sites are enhanced by IKKB-mediated serine phosphorylation at Ser381 20,21 .…”

Section: Introductionmentioning

confidence: 99%

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A zebrafish functional genomics model to investigate the role of human A20 variantsin vivo

Cultrone

Zammit

Self

et al. 2020

Preprint

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24Germline loss-of-function variation in TNFAIP3, encoding A20, has been implicated in a wide 25 variety of autoinflammatory and autoimmune conditions, with acquired somatic missense 26 mutations linked to cancer progression. Furthermore, human sequence data reveals that the A20 27 locus contains ~400 non-synonymous coding variants which are largely uncharacterised. The 28 growing number of A20 coding variants with unknown function, but potential clinical impact, 29 poses a challenge to traditional mouse-based approaches. Here we report the development of a 30 novel functional genomics approach that utilises the new A20-deficient zebrafish (Danio rerio) 31 model to investigate the impact of TNFAIP3 genetic variants in vivo. Similar to A20-deficient 32 mice, A20-deficient zebrafish are hyper-responsive to inflammatory triggers and exhibit 33 spontaneous early lethality. While ectopic addition of human A20 rescued A20-null zebrafish from 34 lethality, missense mutations at two conserved A20 residues, S381A and C243Y reversed this 35 protective effect. Ser381 represents a phosphorylation site important for enhancing A20 activity 36 that is abrogated by its mutation to alanine, or by a C243Y mutation that associates with human 37 autoimmune disease. These data reveal an evolutionarily conserved role for A20, but also 38 demonstrate how a zebrafish functional genomics pipeline can be utilized to investigate the in vivo 39 significance of medically relevant TNFAIP3 gene variants. This approach could be utilised to 40 investigate genetic variation for other conserved genes. 41 42 45activation 2-6 . The medical importance of A20's role in dampening NF-κB is highlighted by cases 46 of germline A20 loss-of-function mutations in humans who present with severe autoinflammatory 47 disease 7,8 , and by the impact of A20 deletion in mice which results in spontaneous and widespread 48 NF-κB activation, multi-organ inflammation and premature lethality 1,9 . The emergence of A20 49 mutations and coding variants as causal genetic factors driving human disease 10,11 highlight the 50 need to better understand A20's functional domains controlling inflammation in vivo. 51 52 A20 suppresses NF-κB signalling with inhibitory activity against key molecular substrates 53 including signalling molecules TNF receptor-associated factor 6 (TRAF6) 9 , receptor interacting 54 protein 1 (RIP1) 12,13 , and the IκB kinase complex (IKK) 14 . The A20 ovarian-tumour (OTU) 55 domain exhibits deubiquitinating editing (DUB) activity cantered on Cys103, which cleaves 56 activating K63-linked ubiquitin chains from RIPK1, TRAF6 and NEMO to terminate NF-κB 57 signalling 9,12,13,15 . The A20 zinc finger 4 (ZnF4) exhibits E3 ligase activity, adding K48-linked 58 ubiquitin chains to RIPK1, triggering RIPK1 proteolysis 12,16 . In addition, the C-terminal ZnF7 59 domain of A20 binds linear ubiquitin to non-catalytically suppress NF-κB activity 17 . A20 is 60 regulated at the level of gene transcription 6 , whereby NF-κB activation induces TNFAIP3 61 expressi...

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“…In addition, A20‐deleted T cells showed increased sensitivity to tumor antigens and antitumor effect (Giordano et al, 2014). Moreover, targeting A20 enhances apoptosis and/or necroptosis in tumor cells and macrophages (Hjelmeland et al, 2010; E. Lee et al, 2019; Polykratis et al, 2019; Zheng et al, 2017). The strategy of targeting A20 is shown in Figure 2.…”

Section: Targeting the Tumor Microenvironment Through Downregulation mentioning

confidence: 99%

In situ antitumor vaccination: Targeting the tumor microenvironment

et al. 2020

Journal Cellular Physiology

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Tumor microenvironment is known to play important roles in tumor progression. Many therapies, targeting the tumor microenvironment, are designed and applied in the clinic. One of these approaches is in situ antitumor therapy. This way, bacteria, antibodies, plasmid DNA, viruses, and cells are intratumorally delivered into the tumor site as “in‐situ antitumor vaccine,” which seeks to enhance immunogenicity and generate systemic T cell responses. In addition, this intratumoral therapy can alter the tumor microenvironment from immunosuppressive to immunostimulatory while limiting the risk of systemic exposure and associated toxicity. Contemporarily, promising preclinical results and some initial success in clinical trials have been obtained after intratumoral therapy.

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A20 prevents inflammasome-dependent arthritis by inhibiting macrophage necroptosis through its ZnF7 ubiquitin-binding domain

Cited by 124 publications

References 64 publications

Beyond inflammasomes: emerging function of gasdermins during apoptosis and NETosis

Beyond inflammasomes: emerging function of gasdermins during apoptosis and NETosis

A zebrafish functional genomics model to investigate the role of human A20 variantsin vivo

In situ antitumor vaccination: Targeting the tumor microenvironment

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