89 results on '"Rehwinkel, Jan"'
Search Results
2. Hypoxia drives HIF2-dependent reversible macrophage cell cycle entry.
- Author
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Meng B, Zhao N, Mlcochova P, Ferreira IATM, Ortmann BM, Davis T, Wit N, Rehwinkel J, Cook S, Maxwell PH, Nathan JA, and Gupta RK
- Subjects
- Animals, Humans, Mice, Mice, Inbred C57BL, Tumor-Associated Macrophages metabolism, Macrophages metabolism, Basic Helix-Loop-Helix Transcription Factors metabolism, Cell Cycle, Cell Hypoxia
- Abstract
Low-oxygen conditions (hypoxia) have been associated primarily with cell-cycle arrest in dividing cells. Macrophages are typically quiescent in G0 but can proliferate in response to tissue signals. Here we show that hypoxia (1% oxygen tension) results in reversible entry into the cell cycle in macrophages. Cell cycle progression is largely limited to G0-G1/S phase transition with little progression to G2/M. This cell cycle transitioning is triggered by an HIF2α-directed transcriptional program. The response is accompanied by increased expression of cell-cycle-associated proteins, including CDK1, which is known to phosphorylate SAMHD1 at T592 and thereby regulate antiviral activity. Prolyl hydroxylase (PHD) inhibitors are able to recapitulate HIF2α-dependent cell cycle entry in macrophages. Finally, tumor-associated macrophages (TAMs) in lung cancers exhibit transcriptomic profiles representing responses to low oxygen and cell cycle progression at the single-cell level. These findings have implications for inflammation and tumor progression/metastasis where low-oxygen environments are common., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
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- 2024
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3. ADAR1: from basic mechanisms to inhibitors.
- Author
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Rehwinkel J and Mehdipour P
- Abstract
Adenosine deaminase acting on RNA 1 (ADAR1) converts adenosine to inosine in double-stranded RNA (dsRNA) molecules, a process known as A-to-I editing. ADAR1 deficiency in humans and mice results in profound inflammatory diseases characterised by the spontaneous induction of innate immunity. In cells lacking ADAR1, unedited RNAs activate RNA sensors. These include melanoma differentiation-associated gene 5 (MDA5) that induces the expression of cytokines, particularly type I interferons (IFNs), protein kinase R (PKR), oligoadenylate synthase (OAS), and Z-DNA/RNA binding protein 1 (ZBP1). Immunogenic RNAs 'defused' by ADAR1 may include transcripts from repetitive elements and other long duplex RNAs. Here, we review these recent fundamental discoveries and discuss implications for human diseases. Some tumours depend on ADAR1 to escape immune surveillance, opening the possibility of unleashing anticancer therapies with ADAR1 inhibitors., Competing Interests: Declaration of interests J.R. is a member of the scientific advisory board and paid consultant for Avammune Therapeutics. P.M. declares no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)
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- 2024
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4. HSV-1 employs UL56 to antagonize expression and function of cGAMP channels.
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Blest HTW, Redmond A, Avissar J, Barker J, Bridgeman A, Fowler G, Chauveau L, Hertzog J, Vendrell I, Fischer R, Iversen MB, Jing L, Koelle DM, Paludan SR, Kessler BM, Crump CM, and Rehwinkel J
- Subjects
- Animals, Humans, HEK293 Cells, Herpes Simplex virology, Herpes Simplex metabolism, Herpes Simplex immunology, Viral Proteins metabolism, Herpesvirus 1, Human physiology, Nucleotides, Cyclic metabolism
- Abstract
DNA sensing is important for antiviral immunity. The DNA sensor cGAS synthesizes 2'3'-cyclic GMP-AMP (cGAMP), a second messenger that activates STING, which induces innate immunity. cGAMP not only activates STING in the cell where it is produced but cGAMP also transfers to other cells. Transporters, channels, and pores (including SLC19A1, SLC46A2, P2X7, ABCC1, and volume-regulated anion channels (VRACs)) release cGAMP into the extracellular space and/or import cGAMP. We report that infection with multiple human viruses depletes some of these cGAMP conduits. This includes herpes simplex virus 1 (HSV-1) that targets SLC46A2, P2X7, and the VRAC subunits LRRC8A and LRRC8C for degradation. The HSV-1 protein UL56 is necessary and sufficient for these effects that are mediated at least partially by proteasomal turnover. UL56 thereby inhibits cGAMP uptake via VRAC, SLC46A2, and P2X7. Taken together, HSV-1 antagonizes intercellular cGAMP transfer. We propose that this limits innate immunity by reducing cell-to-cell communication via the immunotransmitter cGAMP., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2024
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5. ZBP1 activation triggers hematopoietic stem and progenitor cell death resulting in bone marrow failure in mice.
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Roderick-Richardson JE, Lim SE, Suzuki S, Ahmad MH, Selway J, Suleiman R, Karna K, Lehman J, O'Donnell J, Castilla LH, Maelfait J, Rehwinkel J, and Kelliher MA
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- Animals, Humans, Mice, Apoptosis genetics, Bone Marrow Failure Disorders, Cell Death physiology, Hematopoietic Stem Cells metabolism, Necrosis metabolism, Receptor-Interacting Protein Serine-Threonine Kinases metabolism, Nucleic Acids metabolism, Pancytopenia
- Abstract
Human bone marrow failure (BMF) syndromes result from the loss of hematopoietic stem and progenitor cells (HSPC), and this loss has been attributed to cell death; however, the cell death triggers, and mechanisms remain unknown. During BMF, tumor necrosis factor-α (TNFα) and interferon-γ (IFNγ) increase. These ligands are known to induce necroptosis, an inflammatory form of cell death mediated by RIPK1, RIPK3, and MLKL. We previously discovered that mice with a hematopoietic RIPK1 deficiency ( Ripk1
HEM KO ) exhibit inflammation, HSPC loss, and BMF, which is partially ameliorated by a RIPK3 deficiency; however, whether RIPK3 exerts its effects through its function in mediating necroptosis or other forms of cell death remains unclear. Here, we demonstrate that similar to a RIPK3 deficiency, an MLKL deficiency significantly extends survival and like Ripk3 deficiency partially restores hematopoiesis in Ripk1HEM KO mice revealing that both necroptosis and apoptosis contribute to BMF in these mice. Using mouse models, we show that the nucleic acid sensor Z-DNA binding protein 1 (ZBP1) is up-regulated in mouse RIPK1-deficient bone marrow cells and that ZBP1's function in endogenous nucleic acid sensing is necessary for HSPC death and contributes to BMF. We also provide evidence that IFNγ mediates HSPC death in Ripk1HEM KO mice, as ablation of IFNγ but not TNFα receptor signaling significantly extends survival of these mice. Together, these data suggest that RIPK1 maintains hematopoietic homeostasis by preventing ZBP1 activation and induction of HSPC death., Competing Interests: Competing interests statement:The authors declare no competing interest.- Published
- 2024
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6. PARP14 is a PARP with both ADP-ribosyl transferase and hydrolase activities.
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Đukić N, Strømland Ø, Elsborg JD, Munnur D, Zhu K, Schuller M, Chatrin C, Kar P, Duma L, Suyari O, Rack JGM, Baretić D, Crudgington DRK, Groslambert J, Fowler G, Wijngaarden S, Prokhorova E, Rehwinkel J, Schüler H, Filippov DV, Sanyal S, Ahel D, Nielsen ML, Smith R, and Ahel I
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- Humans, Poly(ADP-ribose) Polymerase Inhibitors, Antiviral Agents, Hydrolases, Poly(ADP-ribose) Polymerases genetics, Transferases, COVID-19
- Abstract
PARP14 is a mono-ADP-ribosyl transferase involved in the control of immunity, transcription, and DNA replication stress management. However, little is known about the ADP-ribosylation activity of PARP14, including its substrate specificity or how PARP14-dependent ADP-ribosylation is reversed. We show that PARP14 is a dual-function enzyme with both ADP-ribosyl transferase and hydrolase activity acting on both protein and nucleic acid substrates. In particular, we show that the PARP14 macrodomain 1 is an active ADP-ribosyl hydrolase. We also demonstrate hydrolytic activity for the first macrodomain of PARP9. We reveal that expression of a PARP14 mutant with the inactivated macrodomain 1 results in a marked increase in mono(ADP-ribosyl)ation of proteins in human cells, including PARP14 itself and antiviral PARP13, and displays specific cellular phenotypes. Moreover, we demonstrate that the closely related hydrolytically active macrodomain of SARS2 Nsp3, Mac1, efficiently reverses PARP14 ADP-ribosylation in vitro and in cells, supporting the evolution of viral macrodomains to counteract PARP14-mediated antiviral response.
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- 2023
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7. The Z-nucleic acid sensor ZBP1 in health and disease.
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Maelfait J and Rehwinkel J
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- RNA-Binding Proteins metabolism, Cell Death, RNA metabolism, Nucleic Acids, DNA, Z-Form
- Abstract
Nucleic acid sensing is a central process in the immune system, with far-reaching roles in antiviral defense, autoinflammation, and cancer. Z-DNA binding protein 1 (ZBP1) is a sensor for double-stranded DNA and RNA helices in the unusual left-handed Z conformation termed Z-DNA and Z-RNA. Recent research established ZBP1 as a key upstream regulator of cell death and proinflammatory signaling. Recognition of Z-DNA/RNA by ZBP1 promotes host resistance to viral infection but can also drive detrimental autoinflammation. Additionally, ZBP1 has interesting roles in cancer and other disease settings and is emerging as an attractive target for therapy., (© 2023 Maelfait and Rehwinkel.)
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- 2023
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8. 2022 EULAR points to consider for the measurement, reporting and application of IFN-I pathway activation assays in clinical research and practice.
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Rodríguez-Carrio J, Burska A, Conaghan PG, Dik WA, Biesen R, Eloranta ML, Cavalli G, Visser M, Boumpas DT, Bertsias G, Wahren-Herlenius M, Rehwinkel J, Frémond ML, Crow MK, Rönnblom L, Versnel MA, and Vital EM
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- Humans, Musculoskeletal Diseases, Rheumatology
- Abstract
Background: Type I interferons (IFN-Is) play a role in a broad range of rheumatic and musculoskeletal diseases (RMDs), and compelling evidence suggests that their measurement could have clinical value, although testing has not progressed into clinical settings., Objective: To develop evidence-based points to consider (PtC) for the measurement and reporting of IFN-I assays in clinical research and to determine their potential clinical utility., Methods: EULAR standardised operating procedures were followed. A task force including rheumatologists, immunologists, translational scientists and a patient partner was formed. Two systematic reviews were conducted to address methodological and clinical questions. PtC were formulated based on the retrieved evidence and expert opinion. Level of evidence and agreement was determined., Results: Two overarching principles and 11 PtC were defined. The first set (PtC 1-4) concerned terminology, assay characteristics and reporting practices to enable more consistent reporting and facilitate translation and collaborations. The second set (PtC 5-11) addressed clinical applications for diagnosis and outcome assessments, including disease activity, prognosis and prediction of treatment response. The mean level of agreement was generally high, mainly in the first PtC set and for clinical applications in systemic lupus erythematosus. Harmonisation of assay methodology and clinical validation were key points for the research agenda., Conclusions: IFN-I assays have a high potential for implementation in the clinical management of RMDs. Uptake of these PtC will facilitate the progress of IFN-I assays into clinical practice and may be also of interest beyond rheumatology., Competing Interests: Competing interests: MKC has received consulting fees from AstraZeneca, Bristol Meyers Squibb, Lilly, and Shannon Pharmaceuticals, as well as grant/research support from Gilead. LR has received consulting fees from AstraZeneca. EMV served in the speakers’ bureau of GSK, received consulting fees from AURINIA, SANDOZ, GSK, AstraZeneca, Roche, and Modus, as well as grant/research support from AstraZeneca. PGC has received consultancies or speaker fees from AbbVie, Amgen, AstraZeneca, BMS, Eli Lilly, Galapagos, GSK, Merck, Pfizer, Novartis and UCB., (© Author(s) (or their employer(s)) 2023. No commercial re-use. See rights and permissions. Published by BMJ.)
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- 2023
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9. Association between type I interferon pathway activation and clinical outcomes in rheumatic and musculoskeletal diseases: a systematic literature review informing EULAR points to consider.
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Rodríguez-Carrio J, Burska A, Conaghan PG, Dik WA, Biesen R, Eloranta ML, Cavalli G, Visser M, Boumpas DT, Bertsias G, Wahren-Herlenius M, Rehwinkel J, Frémond ML, Crow MK, Ronnblom L, Vital E, and Versnel M
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- Humans, Interferon Type I, Musculoskeletal Diseases diagnosis, Musculoskeletal Diseases etiology, Myositis, Lupus Erythematosus, Systemic diagnosis
- Abstract
Background: Type I interferons (IFN-I) contribute to a broad range of rheumatic and musculoskeletal diseases (RMDs). Compelling evidence suggests that the measurement of IFN-I pathway activation may have clinical value. Although several IFN-I pathway assays have been proposed, the exact clinical applications are unclear. We summarise the evidence on the potential clinical utility of assays measuring IFN-I pathway activation., Methods: A systematic literature review was conducted across three databases to evaluate the use of IFN-I assays in diagnosis and monitor disease activity, prognosis, response to treatment and responsiveness to change in several RMDs., Results: Of 366 screened, 276 studies were selected that reported the use of assays reflecting IFN-I pathway activation for disease diagnosis (n=188), assessment of disease activity (n=122), prognosis (n=20), response to treatment (n=23) and assay responsiveness (n=59). Immunoassays, quantitative PCR (qPCR) and microarrays were reported most frequently, while systemic lupus erythematosus (SLE), rheumatoid arthritis, myositis, systemic sclerosis and primary Sjögren's syndrome were the most studied RMDs. The literature demonstrated significant heterogeneity in techniques, analytical conditions, risk of bias and application in diseases. Inadequate study designs and technical heterogeneity were the main limitations. IFN-I pathway activation was associated with disease activity and flare occurrence in SLE, but their incremental value was uncertain. IFN-I pathway activation may predict response to IFN-I targeting therapies and may predict response to different treatments., Conclusions: Evidence indicates potential clinical value of assays measuring IFN-I pathway activation in several RMDs, but assay harmonisation and clinical validation are urged. This review informs the EULAR points to consider for the measurement and reporting of IFN-I pathway assays., Competing Interests: Competing interests: MKC has received consulting fees from AstraZeneca, Bristol Meyers Squibb, Lilly and Shannon Pharmaceuticals, as well as grant/research support from Gilead. LR has received consulting fees from AstraZeneca. EV served in the speakers’ bureau of GSK, received consulting fees from AURINIA, SANDOZ, GSK, AstraZeneca, Roche and Modus, as well as grant/research support from AstraZeneca. PGC has received consultancies or speaker fees from AbbVie, Amgen, AstraZeneca, BMS, Eli Lilly, Galapagos, GSK, Merck, Pfizer, Novartis and UCB., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY. Published by BMJ.)
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- 2023
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10. Type I interferon pathway assays in studies of rheumatic and musculoskeletal diseases: a systematic literature review informing EULAR points to consider.
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Burska A, Rodríguez-Carrio J, Biesen R, Dik WA, Eloranta ML, Cavalli G, Visser M, Boumpas DT, Bertsias G, Wahren-Herlenius M, Rehwinkel J, Frémond ML, Crow MK, Ronnblom L, Conaghan PG, Versnel M, and Vital E
- Subjects
- Humans, Reproducibility of Results, Advisory Committees, Interferon Type I, Musculoskeletal Diseases diagnosis, Musculoskeletal Diseases etiology, Rheumatic Diseases diagnosis
- Abstract
Objectives: To systematically review the literature for assay methods that aim to evaluate type I interferon (IFN-I) pathway activation and to harmonise-related terminology., Methods: Three databases were searched for reports of IFN-I and rheumatic musculoskeletal diseases. Information about the performance metrics of assays measuring IFN-I and measures of truth were extracted and summarised. A EULAR task force panel assessed feasibility and developed consensus terminology., Results: Of 10 037 abstracts, 276 fulfilled eligibility criteria for data extraction. Some reported more than one technique to measure IFN-I pathway activation. Hence, 276 papers generated data on 412 methods. IFN-I pathway activation was measured using: qPCR (n=121), immunoassays (n=101), microarray (n=69), reporter cell assay (n=38), DNA methylation (n=14), flow cytometry (n=14), cytopathic effect assay (n=11), RNA sequencing (n=9), plaque reduction assay (n=8), Nanostring (n=5), bisulphite sequencing (n=3). Principles of each assay are summarised for content validity. Concurrent validity (correlation with other IFN assays) was presented for n=150/412 assays. Reliability data were variable and provided for 13 assays. Gene expression and immunoassays were considered most feasible. Consensus terminology to define different aspects of IFN-I research and practice was produced., Conclusions: Diverse methods have been reported as IFN-I assays and these differ in what elements or aspects of IFN-I pathway activation they measure and how. No 'gold standard' represents the entirety of the IFN pathway, some may not be specific for IFN-I. Data on reliability or comparing assays were limited, and feasibility is a challenge for many assays. Consensus terminology should improve consistency of reporting., Competing Interests: Competing interests: MKC has received consulting fees from AstraZeneca, Bristol Meyers Squibb, Lilly and Shannon Pharmaceuticals, as well as grant/research support from Gilead. LR has received consulting fees from AstraZeneca. EV served in the speakers’ bureau of GSK, received consulting fees from AURINIA, SANDOZ, GSK, AstraZeneca, Roche, and Modus, as well as grant/research support from AstraZeneca. PGC has received consultancies or speaker fees from AbbVie, Amgen, AstraZeneca, BMS, Eli Lilly, Galapagos, GSK, Merck, Pfizer, Novartis and UCB. The rest of the authors have no conflict of interest to declare., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY. Published by BMJ.)
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- 2023
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11. A simple transwell-based infection system for obtaining pure populations of VZV-infected cells.
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Hertzog J and Rehwinkel J
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- Humans, Herpesvirus 3, Human, Herpes Zoster, Chickenpox
- Abstract
Varicella-Zoster virus (VZV) is a human herpesvirus and causes chickenpox and shingles. Research into its molecular virology has been hampered by a lack of methods for generation of high-titre, cell-free infectious virus preparations. VZV propagation and infection in vitro are therefore commonly achieved by co-culture of uninfected 'target' cells with infected 'inoculum' cells. A major drawback of this approach is that it results in mixed cell populations after infection. To overcome this limitation we developed a transwell-based VZV infection system. Infected inoculum cells and uninfected target cells are spatially separated by a transwell membrane. While cell-cell contact and VZV spread can occur through membrane pores, the two cell populations do not mix. This simple protocol requires no special instrumentation or reagents. We successfully used this system for infection of a range of target cells and obtained pure populations for downstream analyses such as flow cytometry and RT-qPCR. In sum, we developed a broadly applicable approach to study the molecular and cellular biology as well as host-pathogen interactions of VZV., Competing Interests: Declaration of Interest Statement The authors declare no conflict of interest., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)
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- 2023
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12. Human ZBP1 induces cell death-independent inflammatory signaling via RIPK3 and RIPK1.
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Peng R, Wang CK, Wang-Kan X, Idorn M, Kjaer M, Zhou FY, Fiil BK, Timmermann F, Orozco SL, McCarthy J, Leung CS, Lu X, Bagola K, Rehwinkel J, Oberst A, Maelfait J, Paludan SR, and Gyrd-Hansen M
- Subjects
- Humans, Cytokines, Signal Transduction, Ubiquitin, HT29 Cells, Inflammation, Cell Death, Receptor-Interacting Protein Serine-Threonine Kinases genetics, RNA-Binding Proteins genetics
- Abstract
ZBP1 is an interferon-induced cytosolic nucleic acid sensor that facilitates antiviral responses via RIPK3. Although ZBP1-mediated programmed cell death is widely described, whether and how it promotes inflammatory signaling is unclear. Here, we report a ZBP1-induced inflammatory signaling pathway mediated by K63- and M1-linked ubiquitin chains, which depends on RIPK1 and RIPK3 as scaffolds independently of cell death. In human HT29 cells, ZBP1 associated with RIPK1 and RIPK3 as well as ubiquitin ligases cIAP1 and LUBAC. ZBP1-induced K63- and M1-linked ubiquitination of RIPK1 and ZBP1 to promote TAK1- and IKK-mediated inflammatory signaling and cytokine production. Inhibition of caspase activity suppressed ZBP1-induced cell death but enhanced cytokine production in a RIPK1- and RIPK3 kinase activity-dependent manner. Lastly, we provide evidence that ZBP1 signaling contributes to SARS-CoV-2-induced cytokine production. Taken together, we describe a ZBP1-RIPK3-RIPK1-mediated inflammatory signaling pathway relayed by the scaffolding role of RIPKs and regulated by caspases, which may induce inflammation when ZBP1 is activated below the threshold needed to trigger a cell death response., (© 2022 The Authors. Published under the terms of the CC BY 4.0 license.)
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- 2022
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13. Low expression of EXOSC2 protects against clinical COVID-19 and impedes SARS-CoV-2 replication.
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Moll T, Odon V, Harvey C, Collins MO, Peden A, Franklin J, Graves E, Marshall JN, Dos Santos Souza C, Zhang S, Castelli L, Hautbergue G, Azzouz M, Gordon D, Krogan N, Ferraiuolo L, Snyder MP, Shaw PJ, Rehwinkel J, and Cooper-Knock J
- Subjects
- Chromatography, Liquid, Codon, Nonsense, DNA-Directed RNA Polymerases genetics, Exosome Multienzyme Ribonuclease Complex genetics, Genome-Wide Association Study, Humans, RNA, Viral metabolism, RNA-Binding Proteins genetics, Tandem Mass Spectrometry, Viral Replicase Complex Proteins, Virus Replication genetics, COVID-19 genetics, SARS-CoV-2 genetics
- Abstract
New therapeutic targets are a valuable resource for treatment of SARS-CoV-2 viral infection. Genome-wide association studies have identified risk loci associated with COVID-19, but many loci are associated with comorbidities and are not specific to host-virus interactions. Here, we identify and experimentally validate a link between reduced expression of EXOSC2 and reduced SARS-CoV-2 replication. EXOSC2 was one of the 332 host proteins examined, all of which interact directly with SARS-CoV-2 proteins. Aggregating COVID-19 genome-wide association studies statistics for gene-specific eQTLs revealed an association between increased expression of EXOSC2 and higher risk of clinical COVID-19. EXOSC2 interacts with Nsp8 which forms part of the viral RNA polymerase. EXOSC2 is a component of the RNA exosome, and here, LC-MS/MS analysis of protein pulldowns demonstrated interaction between the SARS-CoV-2 RNA polymerase and most of the human RNA exosome components. CRISPR/Cas9 introduction of nonsense mutations within EXOSC2 in Calu-3 cells reduced EXOSC2 protein expression and impeded SARS-CoV-2 replication without impacting cellular viability. Targeted depletion of EXOSC2 may be a safe and effective strategy to protect against clinical COVID-19., (© 2022 Moll et al.)
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- 2022
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14. Varicella-Zoster virus ORF9 is an antagonist of the DNA sensor cGAS.
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Hertzog J, Zhou W, Fowler G, Rigby RE, Bridgeman A, Blest HT, Cursi C, Chauveau L, Davenne T, Warner BE, Kinchington PR, Kranzusch PJ, and Rehwinkel J
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- DNA metabolism, Humans, Immunity, Innate, Membrane Proteins immunology, Herpesvirus 3, Human genetics, Herpesvirus 3, Human immunology, Interferon Type I immunology, Nucleotidyltransferases antagonists & inhibitors, Nucleotidyltransferases immunology, Viral Proteins immunology
- Abstract
Varicella-Zoster virus (VZV) causes chickenpox and shingles. Although the infection is associated with severe morbidity in some individuals, molecular mechanisms that determine innate immune responses remain poorly defined. We found that the cGAS/STING DNA sensing pathway was required for type I interferon (IFN) induction during VZV infection and that recognition of VZV by cGAS restricted its replication. Screening of a VZV ORF expression library identified the essential VZV tegument protein ORF9 as a cGAS antagonist. Ectopically or virally expressed ORF9 bound to endogenous cGAS leading to reduced type I IFN responses to transfected DNA. Confocal microscopy revealed co-localisation of cGAS and ORF9. ORF9 and cGAS also interacted directly in a cell-free system and phase-separated together with DNA. Furthermore, ORF9 inhibited cGAMP production by cGAS. Taken together, these results reveal the importance of the cGAS/STING DNA sensing pathway for VZV recognition and identify a VZV immune antagonist that partially but directly interferes with DNA sensing via cGAS., (© 2022 The Authors. Published under the terms of the CC BY 4.0 license.)
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- 2022
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15. The interferon-inducible GTPase MxB promotes capsid disassembly and genome release of herpesviruses.
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Serrero MC, Girault V, Weigang S, Greco TM, Ramos-Nascimento A, Anderson F, Piras A, Hickford Martinez A, Hertzog J, Binz A, Pohlmann A, Prank U, Rehwinkel J, Bauerfeind R, Cristea IM, Pichlmair A, Kochs G, and Sodeik B
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- Capsid Proteins metabolism, GTP Phosphohydrolases metabolism, Interferons metabolism, Simplexvirus, Capsid metabolism, Herpesviridae
- Abstract
Host proteins sense viral products and induce defence mechanisms, particularly in immune cells. Using cell-free assays and quantitative mass spectrometry, we determined the interactome of capsid-host protein complexes of herpes simplex virus and identified the large dynamin-like GTPase myxovirus resistance protein B (MxB) as an interferon-inducible protein interacting with capsids. Electron microscopy analyses showed that cytosols containing MxB had the remarkable capability to disassemble the icosahedral capsids of herpes simplex viruses and varicella zoster virus into flat sheets of connected triangular faces. In contrast, capsids remained intact in cytosols with MxB mutants unable to hydrolyse GTP or to dimerize. Our data suggest that MxB senses herpesviral capsids, mediates their disassembly, and thereby restricts the efficiency of nuclear targeting of incoming capsids and/or the assembly of progeny capsids. The resulting premature release of viral genomes from capsids may enhance the activation of DNA sensors, and thereby amplify the innate immune responses., Competing Interests: MS, VG, SW, TG, AR, FA, AP, AH, JH, AB, AP, UP, JR, RB, IC, AP, GK, BS No competing interests declared, (© 2022, Serrero et al.)
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- 2022
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16. Low expression of EXOSC2 protects against clinical COVID-19 and impedes SARS-CoV-2 replication.
- Author
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Moll T, Odon V, Harvey C, Collins MO, Peden A, Franklin J, Graves E, Marshall JNG, Souza CDS, Zhang S, Azzouz M, Gordon D, Krogan N, Ferraiuolo L, Snyder MP, Shaw PJ, Rehwinkel J, and Cooper-Knock J
- Abstract
New therapeutic targets are a valuable resource in the struggle to reduce the morbidity and mortality associated with the COVID-19 pandemic, caused by the SARS-CoV-2 virus. Genome-wide association studies (GWAS) have identified risk loci, but some loci are associated with co-morbidities and are not specific to host-virus interactions. Here, we identify and experimentally validate a link between reduced expression of EXOSC2 and reduced SARS-CoV-2 replication. EXOSC2 was one of 332 host proteins examined, all of which interact directly with SARS-CoV-2 proteins; EXOSC2 interacts with Nsp8 which forms part of the viral RNA polymerase. Lung-specific eQTLs were identified from GTEx (v7) for each of the 332 host proteins. Aggregating COVID-19 GWAS statistics for gene-specific eQTLs revealed an association between increased expression of EXOSC2 and higher risk of clinical COVID-19 which survived stringent multiple testing correction. EXOSC2 is a component of the RNA exosome and indeed, LC-MS/MS analysis of protein pulldowns demonstrated an interaction between the SARS-CoV-2 RNA polymerase and the majority of human RNA exosome components. CRISPR/Cas9 introduction of nonsense mutations within EXOSC2 in Calu-3 cells reduced EXOSC2 protein expression, impeded SARS-CoV-2 replication and upregulated oligoadenylate synthase ( OAS) genes, which have been linked to a successful immune response against SARS-CoV-2. Reduced EXOSC2 expression did not reduce cellular viability. OAS gene expression changes occurred independent of infection and in the absence of significant upregulation of other interferon-stimulated genes (ISGs). Targeted depletion or functional inhibition of EXOSC2 may be a safe and effective strategy to protect at-risk individuals against clinical COVID-19.
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- 2022
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17. Publisher Correction: Chemotherapy-induced transposable elements activate MDA5 to enhance haematopoietic regeneration.
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Clapes T, Polyzou A, Prater P, Sagar, Morales-Hernández A, Ferrarini MG, Kehrer N, Lefkopoulos S, Bergo V, Hummel B, Obier N, Maticzka D, Bridgeman A, Herman JS, Ilik I, Klaeylé L, Rehwinkel J, McKinney-Freeman S, Backofen R, Akhtar A, Cabezas-Wallscheid N, Sawarkar R, Rebollo R, Grün D, and Trompouki E
- Published
- 2021
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18. Hypoxia Regulates Endogenous Double-Stranded RNA Production via Reduced Mitochondrial DNA Transcription.
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Arnaiz E, Miar A, Dias Junior AG, Prasad N, Schulze U, Waithe D, Nathan JA, Rehwinkel J, and Harris AL
- Abstract
Hypoxia is a common phenomenon in solid tumours strongly linked to the hallmarks of cancer. Hypoxia promotes local immunosuppression and downregulates type I interferon (IFN) expression and signalling, which contribute to the success of many cancer therapies. Double-stranded RNA (dsRNA), transiently generated during mitochondrial transcription, endogenously activates the type I IFN pathway. We report the effects of hypoxia on the generation of mitochondrial dsRNA (mtdsRNA) in breast cancer. We found a significant decrease in dsRNA production in different cell lines under hypoxia. This effect was HIF1α/2α-independent. mtdsRNA was responsible for induction of type I IFN and significantly decreased after hypoxia. Mitochondrially encoded gene expression was downregulated and mtdsRNA bound by the dsRNA-specific J2 antibody was decreased during hypoxia. These findings reveal a new mechanism of hypoxia-induced immunosuppression that could be targeted by hypoxia-activated therapies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Arnaiz, Miar, Dias Junior, Prasad, Schulze, Waithe, Nathan, Rehwinkel and Harris.)
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- 2021
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19. Interferon induction held captive in tumor cells.
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Bridgeman A and Rehwinkel J
- Subjects
- Immunity, Innate, RNA, Signal Transduction, Interferons, Nucleic Acids
- Abstract
Unusual nucleic acids activate innate immunity and may be present in transformed cells. Meng et al. (2021) find that cancer-associated mutations in NF2 turn this tumor suppressor into a potent antagonist of DNA- and RNA-induced innate immune signaling., (Copyright © 2021 Elsevier Inc. All rights reserved.)
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- 2021
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20. Adenosine-to-inosine editing of endogenous Z-form RNA by the deaminase ADAR1 prevents spontaneous MAVS-dependent type I interferon responses.
- Author
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Tang Q, Rigby RE, Young GR, Hvidt AK, Davis T, Tan TK, Bridgeman A, Townsend AR, Kassiotis G, and Rehwinkel J
- Subjects
- Adenosine genetics, Adenosine metabolism, Animals, Autoimmune Diseases of the Nervous System genetics, Autoimmune Diseases of the Nervous System immunology, Inosine genetics, Inosine metabolism, Interferon Type I genetics, Mice, Mutation, Nervous System Malformations genetics, Nervous System Malformations immunology, RNA Editing genetics, RNA, Double-Stranded metabolism, Adaptor Proteins, Signal Transducing immunology, Adenosine Deaminase genetics, Interferon Type I immunology, RNA, Double-Stranded genetics
- Abstract
Nucleic acids are powerful triggers of innate immunity and can adopt the Z-conformation, an unusual left-handed double helix. Here, we studied the biological function(s) of Z-RNA recognition by the adenosine deaminase ADAR1, mutations in which cause Aicardi-Goutières syndrome. Adar1
mZα/mZα mice, bearing two point mutations in the Z-nucleic acid binding (Zα) domain that abolish Z-RNA binding, displayed spontaneous induction of type I interferons (IFNs) in multiple organs, including in the lung, where both stromal and hematopoietic cells showed IFN-stimulated gene (ISG) induction. Lung neutrophils expressed ISGs induced by the transcription factor IRF3, indicating an initiating role for neutrophils in this IFN response. The IFN response in Adar1mZα/mZα mice required the adaptor MAVS, implicating cytosolic RNA sensing. Adenosine-to-inosine changes were enriched in transposable elements and revealed a specific requirement of ADAR1's Zα domain in editing of a subset of RNAs. Thus, endogenous RNAs in Z-conformation have immunostimulatory potential curtailed by ADAR1, with relevance to autoinflammatory disease in humans., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2021
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21. Inclusion of cGAMP within virus-like particle vaccines enhances their immunogenicity.
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Chauveau L, Bridgeman A, Tan TK, Beveridge R, Frost JN, Rijal P, Pedroza-Pacheco I, Partridge T, Gilbert-Jaramillo J, Knight ML, Liu X, Russell RA, Borrow P, Drakesmith H, Townsend AR, and Rehwinkel J
- Subjects
- Animals, Humans, Mice, Nucleotides, Cyclic, SARS-CoV-2, Spike Glycoprotein, Coronavirus, COVID-19, Influenza Vaccines, Vaccines, Virus-Like Particle genetics
- Abstract
Cyclic GMP-AMP (cGAMP) is an immunostimulatory molecule produced by cGAS that activates STING. cGAMP is an adjuvant when administered alongside antigens. cGAMP is also incorporated into enveloped virus particles during budding. Here, we investigate whether inclusion of cGAMP within viral vaccine vectors enhances their immunogenicity. We immunise mice with virus-like particles (VLPs) containing HIV-1 Gag and the vesicular stomatitis virus envelope glycoprotein G (VSV-G). cGAMP loading of VLPs augments CD4 and CD8 T-cell responses. It also increases VLP- and VSV-G-specific antibody titres in a STING-dependent manner and enhances virus neutralisation, accompanied by increased numbers of T follicular helper cells. Vaccination with cGAMP-loaded VLPs containing haemagglutinin induces high titres of influenza A virus neutralising antibodies and confers protection upon virus challenge. This requires cGAMP inclusion within VLPs and is achieved at markedly reduced cGAMP doses. Similarly, cGAMP loading of VLPs containing the SARS-CoV-2 Spike protein enhances Spike-specific antibody titres. cGAMP-loaded VLPs are thus an attractive platform for vaccination., (© 2021 The Authors. Published under the terms of the CC BY 4.0 license.)
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- 2021
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22. Fractional response analysis reveals logarithmic cytokine responses in cellular populations.
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Nienałtowski K, Rigby RE, Walczak J, Zakrzewska KE, Głów E, Rehwinkel J, and Komorowski M
- Subjects
- 3T3 Cells, Animals, Healthy Volunteers, Humans, Interferon Type I immunology, Leukocytes, Mononuclear immunology, Mice, Models, Statistical, Primary Cell Culture, Signal Transduction immunology, Single-Cell Analysis, Software, High-Throughput Screening Assays methods, Interferon Type I metabolism, Leukocytes, Mononuclear metabolism, Models, Immunological
- Abstract
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, quantifies 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 heterogeneous cellular populations are sensitive to fold-changes in the dose, as opposed to additive changes.
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- 2021
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23. Chemotherapy-induced transposable elements activate MDA5 to enhance haematopoietic regeneration.
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Clapes T, Polyzou A, Prater P, Sagar, Morales-Hernández A, Ferrarini MG, Kehrer N, Lefkopoulos S, Bergo V, Hummel B, Obier N, Maticzka D, Bridgeman A, Herman JS, Ilik I, Klaeylé L, Rehwinkel J, McKinney-Freeman S, Backofen R, Akhtar A, Cabezas-Wallscheid N, Sawarkar R, Rebollo R, Grün D, and Trompouki E
- Subjects
- Animals, Chromatin Assembly and Disassembly drug effects, Endogenous Retroviruses genetics, Enzyme Activation, HEK293 Cells, Hematopoietic Stem Cells enzymology, Humans, Interferon-Induced Helicase, IFIH1 genetics, Ligands, Long Interspersed Nucleotide Elements, Mice, Inbred C57BL, Mice, Knockout, Signal Transduction, Mice, Cell Proliferation drug effects, Cellular Senescence drug effects, DNA Transposable Elements, Hematopoiesis drug effects, Hematopoietic Stem Cells drug effects, Interferon-Induced Helicase, IFIH1 metabolism, Myeloablative Agonists pharmacology
- Abstract
Haematopoietic stem cells (HSCs) are normally quiescent, but have evolved mechanisms to respond to stress. Here, we evaluate haematopoietic regeneration induced by chemotherapy. We detect robust chromatin reorganization followed by increased transcription of transposable elements (TEs) during early recovery. TE transcripts bind to and activate the innate immune receptor melanoma differentiation-associated protein 5 (MDA5) that generates an inflammatory response that is necessary for HSCs to exit quiescence. HSCs that lack MDA5 exhibit an impaired inflammatory response after chemotherapy and retain their quiescence, with consequent better long-term repopulation capacity. We show that the overexpression of ERV and LINE superfamily TE copies in wild-type HSCs, but not in Mda5
-/- HSCs, results in their cycling. By contrast, after knockdown of LINE1 family copies, HSCs retain their quiescence. Our results show that TE transcripts act as ligands that activate MDA5 during haematopoietic regeneration, thereby enabling HSCs to mount an inflammatory response necessary for their exit from quiescence., (© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2021
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24. The RNA sensor MDA5 detects SARS-CoV-2 infection.
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Sampaio NG, Chauveau L, Hertzog J, Bridgeman A, Fowler G, Moonen JP, Dupont M, Russell RA, Noerenberg M, and Rehwinkel J
- Subjects
- Cell Line, Humans, Immunity, Innate, RNA immunology, Interferon Lambda, COVID-19 immunology, Interferon Type I immunology, Interferon-Induced Helicase, IFIH1 immunology, Interferons immunology, SARS-CoV-2 immunology
- Abstract
Human cells respond to infection by SARS-CoV-2, the virus that causes COVID-19, by producing cytokines including type I and III interferons (IFNs) and proinflammatory factors such as IL6 and TNF. IFNs can limit SARS-CoV-2 replication but cytokine imbalance contributes to severe COVID-19. We studied how cells detect SARS-CoV-2 infection. We report that the cytosolic RNA sensor MDA5 was required for type I and III IFN induction in the lung cancer cell line Calu-3 upon SARS-CoV-2 infection. Type I and III IFN induction further required MAVS and IRF3. In contrast, induction of IL6 and TNF was independent of the MDA5-MAVS-IRF3 axis in this setting. We further found that SARS-CoV-2 infection inhibited the ability of cells to respond to IFNs. In sum, we identified MDA5 as a cellular sensor for SARS-CoV-2 infection that induced type I and III IFNs.
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- 2021
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25. Multi-Modal Characterization of Monocytes in Idiopathic Pulmonary Fibrosis Reveals a Primed Type I Interferon Immune Phenotype.
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Fraser E, Denney L, Antanaviciute A, Blirando K, Vuppusetty C, Zheng Y, Repapi E, Iotchkova V, Taylor S, Ashley N, St Noble V, Benamore R, Hoyles R, Clelland C, Rastrick JMD, Hardman CS, Alham NK, Rigby RE, Simmons A, Rehwinkel J, and Ho LP
- Subjects
- Case-Control Studies, Cells, Cultured, Chemokine CCL2 blood, Flow Cytometry, Gene Expression Profiling, Humans, Idiopathic Pulmonary Fibrosis genetics, Idiopathic Pulmonary Fibrosis metabolism, Idiopathic Pulmonary Fibrosis pathology, Immunophenotyping, Interferon Type I genetics, Interleukin-6 blood, Lung metabolism, Lung pathology, Macrophage Colony-Stimulating Factor blood, Macrophages immunology, Macrophages metabolism, Monocytes metabolism, Phenotype, Receptors, IgG genetics, Receptors, IgG metabolism, Single-Cell Analysis, Idiopathic Pulmonary Fibrosis immunology, Interferon Type I metabolism, Lung immunology, Monocytes immunology
- Abstract
Idiopathic pulmonary fibrosis (IPF) is the most severe form of chronic lung fibrosis. Circulating monocytes have been implicated in immune pathology in IPF but their phenotype is unknown. In this work, we determined the immune phenotype of monocytes in IPF using multi-colour flow cytometry, RNA sequencing and corresponding serum factors, and mapped the main findings to amount of lung fibrosis and single cell transcriptomic landscape of myeloid cells in IPF lungs. We show that monocytes from IPF patients displayed increased expression of CD64 (FcγR1) which correlated with amount of lung fibrosis, and an amplified type I IFN response ex vivo . These were accompanied by markedly raised CSF-1 levels, IL-6, and CCL-2 in serum of IPF patients. Interrogation of single cell transcriptomic data from human IPF lungs revealed increased proportion of CD64
hi monocytes and "transitional macrophages" with higher expression of CCL-2 and type I IFN genes. Our study shows that monocytes in IPF patients are phenotypically distinct from age-matched controls, with a primed type I IFN pathway that may contribute to driving chronic inflammation and fibrosis. These findings strengthen the potential role of monocytes in the pathogenesis of IPF., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Fraser, Denney, Antanaviciute, Blirando, Vuppusetty, Zheng, Repapi, Iotchkova, Taylor, Ashley, St Noble, Benamore, Hoyles, Clelland, Rastrick, Hardman, Alham, Rigby, Simmons, Rehwinkel and Ho.)- Published
- 2021
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26. Innate immunology in COVID-19-a living review. Part I: viral entry, sensing and evasion.
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Coveney C, Tellier M, Lu F, Maleki-Toyserkani S, Jones R, Bart VMT, Pring E, Alrubayyi A, Richter FC, Scourfield DO, Rehwinkel J, Rodrigues PRS, Davies LC, and Gea-Mallorquí E
- Abstract
The coronavirus infectious disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains a world health concern and can cause severe disease and high mortality in susceptible groups. While vaccines offer a chance to treat disease, prophylactic and anti-viral treatments are still of vital importance, especially in context of the mutative ability of this group of viruses. Therefore, it is essential to elucidate the molecular mechanisms of viral entry, innate sensing and immune evasion of SARS-CoV-2, which control the triggers of the subsequent excessive inflammatory response. Viral evasion strategies directly target anti-viral immunity, counteracting host restriction factors and hijacking signalling pathways to interfere with interferon production. In Part I of this review, we examine SARS-CoV-2 viral entry and the described immune evasion mechanisms to provide a perspective on how the failure in initial viral sensing by infected cells can lead to immune dysregulation causing fatal COVID-19, discussed in Part II., (© The Author(s) 2020. Published by Oxford University Press.)
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- 2020
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27. Regulation and inhibition of the DNA sensor cGAS.
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Hertzog J and Rehwinkel J
- Subjects
- Cytokines, DNA genetics, Humans, Signal Transduction, Membrane Proteins genetics, Nucleotidyltransferases genetics, Nucleotidyltransferases metabolism
- Abstract
Cell-autonomous sensing of nucleic acids is essential for host defence against invading pathogens by inducing antiviral and inflammatory cytokines. cGAS has emerged in recent years as a non-redundant DNA sensor important for detection of many viruses and bacteria. Upon binding to DNA, cGAS synthesises the cyclic dinucleotide 2'3'-cGAMP that binds to the adaptor protein STING and thereby triggers IRF3- and NFκB-dependent transcription. In addition to infection, the pathophysiology of an ever-increasing number of sterile inflammatory conditions in humans involves the recognition of DNA through cGAS. Consequently, the cGAS/STING signalling axis has emerged as an attractive target for pharmacological modulation. However, the development of cGAS and STING inhibitors has just begun and a need for specific and effective compounds persists. In this review, we focus on cGAS and explore how its activation by immunostimulatory DNA is regulated by cellular mechanisms, viral immune modulators and small molecules. We further use our knowledge of cGAS modulation by cells and viruses to conceptualise potential new ways of pharmacological cGAS targeting., (© 2020 The Authors.)
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- 2020
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28. cGAS-mediated induction of type I interferon due to inborn errors of histone pre-mRNA processing.
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Uggenti C, Lepelley A, Depp M, Badrock AP, Rodero MP, El-Daher MT, Rice GI, Dhir S, Wheeler AP, Dhir A, Albawardi W, Frémond ML, Seabra L, Doig J, Blair N, Martin-Niclos MJ, Della Mina E, Rubio-Roldán A, García-Pérez JL, Sproul D, Rehwinkel J, Hertzog J, Boland-Auge A, Olaso R, Deleuze JF, Baruteau J, Brochard K, Buckley J, Cavallera V, Cereda C, De Waele LMH, Dobbie A, Doummar D, Elmslie F, Koch-Hogrebe M, Kumar R, Lamb K, Livingston JH, Majumdar A, Lorenço CM, Orcesi S, Peudenier S, Rostasy K, Salmon CA, Scott C, Tonduti D, Touati G, Valente M, van der Linden H Jr, Van Esch H, Vermelle M, Webb K, Jackson AP, Reijns MAM, Gilbert N, and Crow YJ
- Subjects
- Autoimmune Diseases of the Nervous System genetics, Autoimmune Diseases of the Nervous System immunology, Cell Line, DNA immunology, Gene Expression Regulation genetics, Gene Expression Regulation immunology, HCT116 Cells, HEK293 Cells, Hereditary Autoinflammatory Diseases genetics, Hereditary Autoinflammatory Diseases immunology, Humans, Membrane Proteins metabolism, Nervous System Malformations genetics, Nervous System Malformations immunology, Nucleotides, Cyclic biosynthesis, Nucleotidyltransferases metabolism, Chromatin metabolism, Histones metabolism, Interferon Type I biosynthesis, RNA Precursors metabolism, RNA-Binding Proteins genetics, Ribonucleoprotein, U7 Small Nuclear genetics
- Abstract
Inappropriate stimulation or defective negative regulation of the type I interferon response can lead to autoinflammation. In genetically uncharacterized cases of the type I interferonopathy Aicardi-Goutières syndrome, we identified biallelic mutations in LSM11 and RNU7-1, which encode components of the replication-dependent histone pre-mRNA-processing complex. Mutations were associated with the misprocessing of canonical histone transcripts and a disturbance of linker histone stoichiometry. Additionally, we observed an altered distribution of nuclear cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS) and enhanced interferon signaling mediated by the cGAS-stimulator of interferon genes (STING) pathway in patient-derived fibroblasts. Finally, we established that chromatin without linker histone stimulates cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) production in vitro more efficiently. We conclude that nuclear histones, as key constituents of chromatin, are essential in suppressing the immunogenicity of self-DNA.
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- 2020
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29. Hypoxia Induces Transcriptional and Translational Downregulation of the Type I IFN Pathway in Multiple Cancer Cell Types.
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Miar A, Arnaiz E, Bridges E, Beedie S, Cribbs AP, Downes DJ, Beagrie RA, Rehwinkel J, and Harris AL
- Subjects
- Animals, Breast Neoplasms genetics, Breast Neoplasms metabolism, Breast Neoplasms pathology, Cell Line, Tumor, Down-Regulation, Female, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, Humans, Hypoxia-Inducible Factor 1, alpha Subunit genetics, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, Interferon Regulatory Factor-3 genetics, Interferon Regulatory Factor-3 metabolism, Interferon Type I genetics, Mice, RNA, Messenger, Signal Transduction immunology, Single-Cell Analysis, Xenograft Model Antitumor Assays, Interferon Type I metabolism, Neoplasms genetics, Neoplasms metabolism, Neoplasms pathology, Tumor Hypoxia
- Abstract
Hypoxia is a common phenomenon in solid tumors and is strongly linked to hallmarks of cancer. Recent evidence has shown that hypoxia promotes local immune suppression. Type I IFN supports cytotoxic T lymphocytes by stimulating the maturation of dendritic cells and enhancing their capacity to process and present antigens. However, little is known about the relationship between hypoxia and the type I IFN pathway, which comprises the sensing of double-stranded RNA and DNA (dsRNA/dsDNA) followed by IFNα/β secretion and transcriptional activation of IFN-stimulated genes (ISG). In this study, we determined the effects of hypoxia on the type I IFN pathway in breast cancer and the mechanisms involved. In cancer cell lines and xenograft models, mRNA and protein expressions of the type I IFN pathway were downregulated under hypoxic conditions. This pathway was suppressed at each level of signaling, from the dsRNA sensors RIG-I and MDA5, the adaptor MAVS, transcription factors IRF3, IRF7, and STAT1, and several ISG including RIG-I, IRF7, STAT1, and ADAR-p150. Importantly, IFN secretion was reduced under hypoxic conditions. HIF1α- and HIF2α-mediated regulation of gene expression did not explain most of the effects. However, ATAC-seq data revealed in hypoxia that peaks with STAT1 and IRF3 motifs had decreased accessibility. Collectively, these results indicate that hypoxia leads to an overall downregulation of the type I IFN pathway due to repressed transcription and lower chromatin accessibility in an HIF1/2α-independent manner, which could contribute to immunosuppression in hypoxic tumors. SIGNIFICANCE: These findings characterize a new mechanism of immunosuppression by hypoxia via downregulation of the type I IFN pathway and its autocrine/paracrine effects on tumor growth., (©2020 American Association for Cancer Research.)
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- 2020
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30. Mutations in COPA lead to abnormal trafficking of STING to the Golgi and interferon signaling.
- Author
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Lepelley A, Martin-Niclós MJ, Le Bihan M, Marsh JA, Uggenti C, Rice GI, Bondet V, Duffy D, Hertzog J, Rehwinkel J, Amselem S, Boulisfane-El Khalifi S, Brennan M, Carter E, Chatenoud L, Chhun S, Coulomb l'Hermine A, Depp M, Legendre M, Mackenzie KJ, Marey J, McDougall C, McKenzie KJ, Molina TJ, Neven B, Seabra L, Thumerelle C, Wislez M, Nathan N, Manel N, Crow YJ, and Frémond ML
- Subjects
- Adolescent, Adult, Child, Endoplasmic Reticulum metabolism, Female, Gene Knockout Techniques, HEK293 Cells, Humans, Male, Membrane Proteins genetics, Middle Aged, Protein Transport genetics, THP-1 Cells, Transfection, Young Adult, Coatomer Protein genetics, Coatomer Protein metabolism, Golgi Apparatus metabolism, Interferon Type I metabolism, Membrane Proteins metabolism, Mutation, Missense, Signal Transduction genetics
- Abstract
Heterozygous missense mutations in coatomer protein subunit α, COPA, cause a syndrome overlapping clinically with type I IFN-mediated disease due to gain-of-function in STING, a key adaptor of IFN signaling. Recently, increased levels of IFN-stimulated genes (ISGs) were described in COPA syndrome. However, the link between COPA mutations and IFN signaling is unknown. We observed elevated levels of ISGs and IFN-α in blood of symptomatic COPA patients. In vitro, both overexpression of mutant COPA and silencing of COPA induced STING-dependent IFN signaling. We detected an interaction between COPA and STING, and mutant COPA was associated with an accumulation of ER-resident STING at the Golgi. Given the known role of the coatomer protein complex I, we speculate that loss of COPA function leads to enhanced type I IFN signaling due to a failure of Golgi-to-ER STING retrieval. These data highlight the importance of the ER-Golgi axis in the control of autoinflammation and inform therapeutic strategies in COPA syndrome., Competing Interests: Disclosures: M. Wislez reported personal fees from Boeringher Ingelheim, Roche, MSD, BMS, Astra Zeneca, and Amgen outside the submitted work. Y.J. Crow reported "other" from Biogen outside the submitted work. No other disclosures were reported., (© 2020 Lepelley et al.)
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- 2020
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31. Author Correction: SARS-CoV2-mediated suppression of NRF2-signaling reveals potent antiviral and anti-inflammatory activity of 4-octyl-itaconate and dimethyl fumarate.
- Author
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Olagnier D, Farahani E, Thyrsted J, Blay-Cadanet J, Herengt A, Idorn M, Hait A, Hernaez B, Knudsen A, Iversen MB, Schilling M, Jørgensen SE, Thomsen M, Reinert LS, Lappe M, Hoang HD, Gilchrist VH, Hansen AL, Ottosen R, Nielsen CG, Møller C, van der Horst D, Peri S, Balachandran S, Huang J, Jakobsen M, Svenningsen EB, Poulsen TB, Bartsch L, Thielke AL, Luo Y, Alain T, Rehwinkel J, Alcamí A, Hiscott J, Mogensen TH, Paludan SR, and Holm CK
- Abstract
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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- 2020
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32. SARS-CoV2-mediated suppression of NRF2-signaling reveals potent antiviral and anti-inflammatory activity of 4-octyl-itaconate and dimethyl fumarate.
- Author
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Olagnier D, Farahani E, Thyrsted J, Blay-Cadanet J, Herengt A, Idorn M, Hait A, Hernaez B, Knudsen A, Iversen MB, Schilling M, Jørgensen SE, Thomsen M, Reinert LS, Lappe M, Hoang HD, Gilchrist VH, Hansen AL, Ottosen R, Nielsen CG, Møller C, van der Horst D, Peri S, Balachandran S, Huang J, Jakobsen M, Svenningsen EB, Poulsen TB, Bartsch L, Thielke AL, Luo Y, Alain T, Rehwinkel J, Alcamí A, Hiscott J, Mogensen TH, Paludan SR, and Holm CK
- Subjects
- Adult, Antioxidants pharmacology, Betacoronavirus metabolism, COVID-19, Coronavirus Infections virology, Dimethyl Fumarate pharmacology, Female, Gene Expression, Gene Knockdown Techniques, Humans, Interferon Type I, Lung pathology, Male, NF-E2-Related Factor 2 genetics, Pandemics, Pneumonia, Viral virology, SARS-CoV-2, Signal Transduction drug effects, Succinates pharmacology, Virus Replication drug effects, Anti-Inflammatory Agents pharmacology, Antiviral Agents pharmacology, Betacoronavirus drug effects, Coronavirus Infections drug therapy, Dimethyl Fumarate agonists, NF-E2-Related Factor 2 metabolism, Pneumonia, Viral drug therapy, Succinates agonists
- Abstract
Antiviral strategies to inhibit Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV2) and the pathogenic consequences of COVID-19 are urgently required. Here, we demonstrate that the NRF2 antioxidant gene expression pathway is suppressed in biopsies obtained from COVID-19 patients. Further, we uncover that NRF2 agonists 4-octyl-itaconate (4-OI) and the clinically approved dimethyl fumarate (DMF) induce a cellular antiviral program that potently inhibits replication of SARS-CoV2 across cell lines. The inhibitory effect of 4-OI and DMF extends to the replication of several other pathogenic viruses including Herpes Simplex Virus-1 and-2, Vaccinia virus, and Zika virus through a type I interferon (IFN)-independent mechanism. In addition, 4-OI and DMF limit host inflammatory responses to SARS-CoV2 infection associated with airway COVID-19 pathology. In conclusion, NRF2 agonists 4-OI and DMF induce a distinct IFN-independent antiviral program that is broadly effective in limiting virus replication and in suppressing the pro-inflammatory responses of human pathogenic viruses, including SARS-CoV2.
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- 2020
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33. PNP inhibitors selectively kill cancer cells lacking SAMHD1.
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Davenne T and Rehwinkel J
- Abstract
Purine nucleoside phosphorylase inhibitors (PNP-Is) were developed to ablate transformed lymphocytes. However, only some patients with leukemia benefit from PNP-Is. We provide a molecular explanation: the deoxyribonucleoside triphosphate (dNTP) hydrolase SAM and HD domain-containing protein 1 (SAMHD1) prevents the accumulation of toxic dNTP levels during purine nucleoside phosphorylase inhibition. We propose PNP-Is for targeted therapy of patients with acquired SAMHD1 mutations., (© 2020 The Author(s). Published with license by Taylor & Francis Group, LLC.)
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- 2020
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34. RIG-I-like receptors: their regulation and roles in RNA sensing.
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Rehwinkel J and Gack MU
- Subjects
- Animals, Humans, Immune System physiology, RNA, Viral metabolism, DEAD Box Protein 58 physiology, RNA metabolism
- Abstract
Retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) are key sensors of virus infection, mediating the transcriptional induction of type I interferons and other genes that collectively establish an antiviral host response. Recent studies have revealed that both viral and host-derived RNAs can trigger RLR activation; this can lead to an effective antiviral response but also immunopathology if RLR activities are uncontrolled. In this Review, we discuss recent advances in our understanding of the types of RNA sensed by RLRs in the contexts of viral infection, malignancies and autoimmune diseases. We further describe how the activity of RLRs is controlled by host regulatory mechanisms, including RLR-interacting proteins, post-translational modifications and non-coding RNAs. Finally, we discuss key outstanding questions in the RLR field, including how our knowledge of RLR biology could be translated into new therapeutics.
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- 2020
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35. SAMHD1-mediated dNTP degradation is required for efficient DNA repair during antibody class switch recombination.
- Author
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Husain A, Xu J, Fujii H, Nakata M, Kobayashi M, Wang JY, Rehwinkel J, Honjo T, and Begum NA
- Subjects
- Cell Line, Deoxyribonucleotides genetics, Humans, SAM Domain and HD Domain-Containing Protein 1 genetics, DNA Repair, Deoxyribonucleotides metabolism, Immunoglobulin Class Switching, SAM Domain and HD Domain-Containing Protein 1 metabolism
- Abstract
Sterile alpha motif and histidine-aspartic acid domain-containing protein 1 (SAMHD1), a dNTP triphosphohydrolase, regulates the levels of cellular dNTPs through their hydrolysis. SAMHD1 protects cells from invading viruses that depend on dNTPs to replicate and is frequently mutated in cancers and Aicardi-Goutières syndrome, a hereditary autoimmune encephalopathy. We discovered that SAMHD1 localizes at the immunoglobulin (Ig) switch region, and serves as a novel DNA repair regulator of Ig class switch recombination (CSR). Depletion of SAMHD1 impaired not only CSR but also IgH/c-Myc translocation. Consistently, we could inhibit these two processes by elevating the cellular nucleotide pool. A high frequency of nucleotide insertion at the break-point junctions is a notable feature in SAMHD1 deficiency during activation-induced cytidine deaminase-mediated genomic instability. Interestingly, CSR induced by staggered but not blunt, double-stranded DNA breaks was impaired by SAMHD1 depletion, which was accompanied by enhanced nucleotide insertions at recombination junctions. We propose that SAMHD1-mediated dNTP balance regulates dNTP-sensitive DNA end-processing enzyme and promotes CSR and aberrant genomic rearrangements by suppressing the insertional DNA repair pathway., (© 2020 The Authors.)
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- 2020
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36. Sensing of endogenous nucleic acids by ZBP1 induces keratinocyte necroptosis and skin inflammation.
- Author
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Devos M, Tanghe G, Gilbert B, Dierick E, Verheirstraeten M, Nemegeer J, de Reuver R, Lefebvre S, De Munck J, Rehwinkel J, Vandenabeele P, Declercq W, and Maelfait J
- Subjects
- Animals, HEK293 Cells, Humans, Inflammation immunology, Interleukin-17 metabolism, Mice, Inbred C57BL, Mice, Knockout, Protein Kinases metabolism, Inflammation pathology, Keratinocytes metabolism, Keratinocytes pathology, Necroptosis, Nucleic Acids metabolism, RNA-Binding Proteins metabolism, Skin pathology
- Abstract
Aberrant detection of endogenous nucleic acids by the immune system can cause inflammatory disease. The scaffold function of the signaling kinase RIPK1 limits spontaneous activation of the nucleic acid sensor ZBP1. Consequently, loss of RIPK1 in keratinocytes induces ZBP1-dependent necroptosis and skin inflammation. Whether nucleic acid sensing is required to activate ZBP1 in RIPK1-deficient conditions and which immune pathways are associated with skin disease remained open questions. Using knock-in mice with disrupted ZBP1 nucleic acid-binding activity, we report that sensing of endogenous nucleic acids by ZBP1 is critical in driving skin pathology characterized by antiviral and IL-17 immune responses. Inducing ZBP1 expression by interferons triggers necroptosis in RIPK1-deficient keratinocytes, and epidermis-specific deletion of MLKL prevents disease, demonstrating that cell-intrinsic events cause inflammation. These findings indicate that dysregulated sensing of endogenous nucleic acid by ZBP1 can drive inflammation and may contribute to the pathogenesis of IL-17-driven inflammatory skin conditions such as psoriasis., Competing Interests: Disclosures: The authors declare no competing interests exist., (© 2020 Devos et al.)
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- 2020
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37. Redox homeostasis maintained by GPX4 facilitates STING activation.
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Jia M, Qin D, Zhao C, Chai L, Yu Z, Wang W, Tong L, Lv L, Wang Y, Rehwinkel J, Yu J, and Zhao W
- Subjects
- Animals, Carbolines pharmacology, Cells, Cultured, DNA, Viral immunology, Disease Models, Animal, Endoplasmic Reticulum metabolism, Female, Fibroblasts, Golgi Apparatus metabolism, HEK293 Cells, Herpes Simplex virology, Herpesvirus 1, Human genetics, Herpesvirus 1, Human immunology, Homeostasis immunology, Humans, Immunity, Innate, Lipid Peroxidation genetics, Lipid Peroxidation immunology, Macrophages, Peritoneal cytology, Macrophages, Peritoneal immunology, Macrophages, Peritoneal metabolism, Membrane Proteins immunology, Mice, Mice, Knockout, Nucleotidyltransferases metabolism, Oxidation-Reduction, Oximes pharmacology, Phospholipid Hydroperoxide Glutathione Peroxidase antagonists & inhibitors, Phospholipid Hydroperoxide Glutathione Peroxidase genetics, Primary Cell Culture, Protein Carbonylation immunology, Signal Transduction drug effects, Signal Transduction immunology, Sulfonamides pharmacology, THP-1 Cells, Virus Replication immunology, Herpes Simplex immunology, Membrane Proteins metabolism, Phospholipid Hydroperoxide Glutathione Peroxidase metabolism
- Abstract
Stimulator-of-interferon genes (STING) is vital for sensing cytosolic DNA and initiating innate immune responses against microbial infection and tumors. Redox homeostasis is the balance of oxidative and reducing reactions present in all living systems. Yet, how the intracellular redox state controls STING activation is unclear. Here, we show that cellular redox homeostasis maintained by glutathione peroxidase 4 (GPX4) is required for STING activation. GPX4 deficiency enhanced cellular lipid peroxidation and thus specifically inhibited the cGAS-STING pathway. Concordantly, GPX4 deficiency inhibited herpes simplex virus-1 (HSV-1)-induced innate antiviral immune responses and promoted HSV-1 replication in vivo. Mechanistically, GPX4 inactivation increased production of lipid peroxidation, which led to STING carbonylation at C88 and inhibited its trafficking from the endoplasmic reticulum (ER) to the Golgi complex. Thus, cellular stress-induced lipid peroxidation specifically attenuates the STING DNA-sensing pathway, suggesting that GPX4 facilitates STING activation by maintaining redox homeostasis of lipids.
- Published
- 2020
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38. RIG-I Plays a Dominant Role in the Induction of Transcriptional Changes in Zika Virus-Infected Cells, which Protect from Virus-Induced Cell Death.
- Author
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Schilling M, Bridgeman A, Gray N, Hertzog J, Hublitz P, Kohl A, and Rehwinkel J
- Subjects
- Animals, Chlorocebus aethiops virology, Humans, Immunity, Innate immunology, Signal Transduction immunology, Vero Cells virology, Viral Nonstructural Proteins metabolism, Zika Virus immunology, Zika Virus metabolism, Zika Virus Infection immunology, Cell Death physiology, DEAD Box Protein 58 metabolism, Receptors, Immunologic metabolism, Zika Virus Infection virology
- Abstract
The Zika virus (ZIKV) has received much attention due to an alarming increase in cases of neurological disorders including congenital Zika syndrome associated with infection. To date, there is no effective treatment available. An immediate response by the innate immune system is crucial for effective control of the virus. Using CRISPR/Cas9-mediated knockouts in A549 cells, we investigated the individual contributions of the RIG-I-like receptors MDA5 and RIG-I to ZIKV sensing and control of this virus by using a Brazilian ZIKV strain. We show that RIG-I is the main sensor for ZIKV in A549 cells. Surprisingly, we observed that loss of RIG-I and consecutive type I interferon (IFN) production led to virus-induced apoptosis. ZIKV non-structural protein NS5 was reported to interfere with type I IFN receptor signaling. Additionally, we show that ZIKV NS5 inhibits type I IFN induction. Overall, our study highlights the importance of RIG-I-dependent ZIKV sensing for the prevention of virus-induced cell death and shows that NS5 inhibits the production of type I IFN.
- Published
- 2020
- Full Text
- View/download PDF
39. SAMHD1 Limits the Efficacy of Forodesine in Leukemia by Protecting Cells against the Cytotoxicity of dGTP.
- Author
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Davenne T, Klintman J, Sharma S, Rigby RE, Blest HTW, Cursi C, Bridgeman A, Dadonaite B, De Keersmaecker K, Hillmen P, Chabes A, Schuh A, and Rehwinkel J
- Subjects
- Animals, Drug Resistance, Neoplasm, Female, Humans, Leukemia, Lymphocytic, Chronic, B-Cell drug therapy, Male, Mice, Mice, Inbred C57BL, Deoxyguanine Nucleotides metabolism, Purine Nucleosides pharmacology, Pyrimidinones pharmacology, SAM Domain and HD Domain-Containing Protein 1 metabolism
- Abstract
The anti-leukemia agent forodesine causes cytotoxic overload of intracellular deoxyguanosine triphosphate (dGTP) but is efficacious only in a subset of patients. We report that SAMHD1, a phosphohydrolase degrading deoxyribonucleoside triphosphate (dNTP), protects cells against the effects of dNTP imbalances. SAMHD1-deficient cells induce intrinsic apoptosis upon provision of deoxyribonucleosides, particularly deoxyguanosine (dG). Moreover, dG and forodesine act synergistically to kill cells lacking SAMHD1. Using mass cytometry, we find that these compounds kill SAMHD1-deficient malignant cells in patients with chronic lymphocytic leukemia (CLL). Normal cells and CLL cells from patients without SAMHD1 mutation are unaffected. We therefore propose to use forodesine as a precision medicine for leukemia, stratifying patients by SAMHD1 genotype or expression., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
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40. Enhanced Immunogenicity of Mitochondrial-Localized Proteins in Cancer Cells.
- Author
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Prota G, Gileadi U, Rei M, Lechuga-Vieco AV, Chen JL, Galiani S, Bedard M, Lau VWC, Fanchi LF, Artibani M, Hu Z, Gordon S, Rehwinkel J, Enríquez JA, Ahmed AA, Schumacher TN, and Cerundolo V
- Subjects
- Animals, Cell Line, Tumor, Disease Models, Animal, Mice, Mice, Inbred C57BL, Mice, Knockout, Mice, Transgenic, Mitochondrial Proteins metabolism, Neoplasms metabolism, Neoplasms therapy, Antigens, Neoplasm immunology, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Cancer Vaccines immunology, Epitopes immunology, Mitochondrial Proteins immunology, Neoplasms immunology
- Abstract
Epitopes derived from mutated cancer proteins elicit strong antitumor T-cell responses that correlate with clinical efficacy in a proportion of patients. However, it remains unclear whether the subcellular localization of mutated proteins influences the efficiency of T-cell priming. To address this question, we compared the immunogenicity of NY-ESO-1 and OVA localized either in the cytosol or in mitochondria. We showed that tumors expressing mitochondrial-localized NY-ESO-1 and OVA proteins elicit significantdly higher frequencies of antigen-specific CD8
+ T cells in vivo . We also demonstrated that this stronger immune response is dependent on the mitochondrial location of the antigenic proteins, which contributes to their higher steady-state amount, compared with cytosolic localized proteins. Consistent with these findings, we showed that injection of mitochondria purified from B16 melanoma cells can protect mice from a challenge with B16 cells, but not with irrelevant tumors. Finally, we extended these findings to cancer patients by demonstrating the presence of T-cell responses specific for mutated mitochondrial-localized proteins. These findings highlight the utility of prioritizing epitopes derived from mitochondrial-localized mutated proteins as targets for cancer vaccination strategies., (©2020 American Association for Cancer Research.)- Published
- 2020
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41. Nucleic Acid Sensors and Programmed Cell Death.
- Author
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Maelfait J, Liverpool L, and Rehwinkel J
- Subjects
- DEAD Box Protein 58 chemistry, DEAD Box Protein 58 genetics, DNA-Binding Proteins chemistry, DNA-Binding Proteins genetics, Humans, Interferon Type I genetics, Interleukin-1beta genetics, Nucleic Acids chemistry, Nucleotidyltransferases chemistry, Nucleotidyltransferases genetics, RNA-Binding Proteins genetics, Receptors, Immunologic, Cell Death genetics, Immunity, Innate genetics, Nucleic Acids genetics
- Abstract
Nucleic acids derived from microorganisms are powerful triggers for innate immune responses. Proteins called RNA and DNA sensors detect foreign nucleic acids and, in mammalian cells, include RIG-I, cGAS, and AIM2. On binding to nucleic acids, these proteins initiate signaling cascades that activate host defense responses. An important aspect of this defense program is the production of cytokines such as type I interferons and IL-1β. Studies conducted over recent years have revealed that nucleic acid sensors also activate programmed cell death pathways as an innate immune response to infection. Indeed, RNA and DNA sensors induce apoptosis, pyroptosis, and necroptosis. Cell death via these pathways prevents replication of pathogens by eliminating the infected cell and additionally contributes to the release of cytokines and inflammatory mediators. Interestingly, recent evidence suggests that programmed cell death triggered by nucleic acid sensors plays an important role in a number of noninfectious pathologies. In addition to nonself DNA and RNA from microorganisms, nucleic acid sensors also recognize endogenous nucleic acids, for example when cells are damaged by genotoxic agents and in certain autoinflammatory diseases. This review article summarizes current knowledge on the links between nucleic acid sensing and cell death and explores important open questions for future studies in this area., (Copyright © 2019 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2020
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42. Deoxyguanosine is a TLR7 agonist.
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Davenne T, Bridgeman A, Rigby RE, and Rehwinkel J
- Subjects
- Animals, Deoxyguanosine metabolism, Humans, Inflammation metabolism, Leukocytes, Mononuclear immunology, Leukocytes, Mononuclear metabolism, Macrophages immunology, Macrophages metabolism, Mice, Mice, Inbred C57BL, Deoxyguanosine immunology, Inflammation immunology, Toll-Like Receptor 7 agonists
- Abstract
Toll-like receptor 7 (TLR7) is an innate immune sensor for single-strand RNA (ssRNA). Recent structural analysis revealed that TLR7 has an additional binding site for nucleosides such as guanosine, and is activated when both guanosine and ssRNA bind. The nucleoside binding site also accommodates imidazoquinoline derivatives such as R848, which activate TLR7 in the absence of ssRNA. Here, we report that deoxyguanosine (dG) triggered cytokine production in murine bone marrow derived macrophages and plasmacytoid dendritic cells, as well as in human peripheral blood mononuclear cells, including type I interferons and pro-inflammatory factors such as TNF and IL-6. This signalling activity of dG was dependent on TLR7 and its adaptor MyD88 and did not require amplification via the type I interferon receptor. dG-triggered cytokine production required endosomal maturation but did not depend on the concurrent provision of RNA. We conclude that dG induces an inflammatory response through TLR7 and propose that dG is an RNA-independent TLR7 agonist., (© 2019 The Authors. European Journal of Immunology published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2020
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43. PA-X antagonises MAVS-dependent accumulation of early type I interferon messenger RNAs during influenza A virus infection.
- Author
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Rigby RE, Wise HM, Smith N, Digard P, and Rehwinkel J
- Subjects
- Adaptor Proteins, Signal Transducing deficiency, Adaptor Proteins, Signal Transducing metabolism, Animals, Immunity, Innate, Influenza A virus metabolism, Interferon Type I genetics, Lung metabolism, Lung virology, Membrane Glycoproteins deficiency, Membrane Glycoproteins genetics, Mice, Mice, Inbred C57BL, Mice, Knockout, Orthomyxoviridae Infections pathology, Orthomyxoviridae Infections virology, RNA, Messenger metabolism, Repressor Proteins deficiency, Repressor Proteins metabolism, Signal Transduction, Toll-Like Receptor 7 deficiency, Toll-Like Receptor 7 genetics, Viral Nonstructural Proteins deficiency, Viral Nonstructural Proteins metabolism, Adaptor Proteins, Signal Transducing genetics, Influenza A virus physiology, Interferon Type I metabolism, Repressor Proteins genetics, Viral Nonstructural Proteins genetics
- Abstract
The sensing of viral nucleic acids by the innate immune system activates a potent antiviral response in the infected cell, a key component of which is the expression of genes encoding type I interferons (IFNs). Many viruses counteract this response by blocking the activation of host nucleic acid sensors. The evolutionarily conserved influenza A virus (IAV) protein PA-X has been implicated in suppressing the host response to infection, including the expression of type I IFNs. Here, we characterise this further using a PA-X-deficient virus of the mouse-adapted PR8 strain to study activation of the innate immune response in a mouse model of the early response to viral infection. We show that levels of Ifna4 and Ifnb1 mRNAs in the lungs of infected mice were elevated in the absence of PA-X and that this was completely dependent on MAVS. This therefore suggests a role for PA-X in preventing the accumulation of early type I IFN mRNAs in the lung during IAV infection.
- Published
- 2019
- Full Text
- View/download PDF
44. A dual role for SAMHD1 in regulating HBV cccDNA and RT-dependent particle genesis.
- Author
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Wing PA, Davenne T, Wettengel J, Lai AG, Zhuang X, Chakraborty A, D'Arienzo V, Kramer C, Ko C, Harris JM, Schreiner S, Higgs M, Roessler S, Parish JL, Protzer U, Balfe P, Rehwinkel J, and McKeating JA
- Subjects
- DNA, Viral genetics, Gene Knockout Techniques, Hep G2 Cells, Hepatitis B, Chronic enzymology, Hepatitis B, Chronic virology, Humans, Reverse Transcription genetics, Transcriptional Activation, Transfection, Virus Replication genetics, DNA, Circular genetics, Hepatitis B virus genetics, RNA-Directed DNA Polymerase genetics, SAM Domain and HD Domain-Containing Protein 1 genetics
- Abstract
Chronic hepatitis B is one of the world's unconquered diseases with more than 240 million infected subjects at risk of developing liver disease and hepatocellular carcinoma. Hepatitis B virus reverse transcribes pre-genomic RNA to relaxed circular DNA (rcDNA) that comprises the infectious particle. To establish infection of a naïve target cell, the newly imported rcDNA is repaired by host enzymes to generate covalently closed circular DNA (cccDNA), which forms the transcriptional template for viral replication. SAMHD1 is a component of the innate immune system that regulates deoxyribonucleoside triphosphate levels required for host and viral DNA synthesis. Here, we show a positive role for SAMHD1 in regulating cccDNA formation, where KO of SAMHD1 significantly reduces cccDNA levels that was reversed by expressing wild-type but not a mutated SAMHD1 lacking the nuclear localization signal. The limited pool of cccDNA in infected Samhd1 KO cells is transcriptionally active, and we observed a 10-fold increase in newly synthesized rcDNA-containing particles, demonstrating a dual role for SAMHD1 to both facilitate cccDNA genesis and to restrict reverse transcriptase-dependent particle genesis., (© 2019 Wing et al.)
- Published
- 2019
- Full Text
- View/download PDF
45. Antiviral activity of bone morphogenetic proteins and activins.
- Author
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Eddowes LA, Al-Hourani K, Ramamurthy N, Frankish J, Baddock HT, Sandor C, Ryan JD, Fusco DN, Arezes J, Giannoulatou E, Boninsegna S, Chevaliez S, Owens BMJ, Sun CC, Fabris P, Giordani MT, Martines D, Vukicevic S, Crowe J, Lin HY, Rehwinkel J, McHugh PJ, Binder M, Babitt JL, Chung RT, Lawless MW, Armitage AE, Webber C, Klenerman P, and Drakesmith H
- Subjects
- Antiviral Agents metabolism, Cells, Cultured, Endopeptidases genetics, Hepacivirus drug effects, Hepatitis C drug therapy, Hepatitis C metabolism, Hepcidins genetics, Humans, Interferon Regulatory Factors genetics, Interferon-alpha pharmacology, Interferon-alpha therapeutic use, RNA, Viral metabolism, Signal Transduction genetics, Smad1 Protein genetics, Ubiquitin Thiolesterase, Virus Replication drug effects, Zika Virus drug effects, Activins pharmacology, Antiviral Agents pharmacology, Bone Morphogenetic Protein 6 pharmacology, Gene Expression Regulation drug effects, Signal Transduction drug effects
- Abstract
Understanding the control of viral infections is of broad importance. Chronic hepatitis C virus (HCV) infection causes decreased expression of the iron hormone hepcidin, which is regulated by hepatic bone morphogenetic protein (BMP)/SMAD signalling. We found that HCV infection and the BMP/SMAD pathway are mutually antagonistic. HCV blunted induction of hepcidin expression by BMP6, probably via tumour necrosis factor (TNF)-mediated downregulation of the BMP co-receptor haemojuvelin. In HCV-infected patients, disruption of the BMP6/hepcidin axis and genetic variation associated with the BMP/SMAD pathway predicted the outcome of infection, suggesting that BMP/SMAD activity influences antiviral immunity. Correspondingly, BMP6 regulated a gene repertoire reminiscent of type I interferon (IFN) signalling, including upregulating interferon regulatory factors (IRFs) and downregulating an inhibitor of IFN signalling, USP18. Moreover, in BMP-stimulated cells, SMAD1 occupied loci across the genome, similar to those bound by IRF1 in IFN-stimulated cells. Functionally, BMP6 enhanced the transcriptional and antiviral response to IFN, but BMP6 and related activin proteins also potently blocked HCV replication independently of IFN. Furthermore, BMP6 and activin A suppressed growth of HBV in cell culture, and activin A inhibited Zika virus replication alone and in combination with IFN. The data establish an unappreciated important role for BMPs and activins in cellular antiviral immunity, which acts independently of, and modulates, IFN.
- Published
- 2019
- Full Text
- View/download PDF
46. A Balancing Act: MDA5 in Antiviral Immunity and Autoinflammation.
- Author
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Dias Junior AG, Sampaio NG, and Rehwinkel J
- Subjects
- Animals, Humans, Interferon-Induced Helicase, IFIH1 genetics, Mutation, Hereditary Autoinflammatory Diseases genetics, Immunity, Innate, Interferon-Induced Helicase, IFIH1 metabolism, RNA, Viral metabolism, Receptors, Immunologic metabolism, Virus Diseases immunology
- Abstract
Induction of interferons during viral infection is mediated by cellular proteins that recognise viral nucleic acids. MDA5 is one such sensor of virus presence and is activated by RNA. MDA5 is required for immunity against several classes of viruses, including picornaviruses. Recent work showed that mutations in the IFIH1 gene, encoding MDA5, lead to interferon-driven autoinflammatory diseases. Together with observations made in cancer cells, this suggests that MDA5 detects cellular RNAs in addition to viral RNAs. It is therefore important to understand the properties of the RNAs which activate MDA5. New data indicate that RNA length and secondary structure are features sensed by MDA5. We review these developments and discuss how MDA5 strikes a balance between antiviral immunity and autoinflammation., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2019
- Full Text
- View/download PDF
47. Nitro-fatty acids are formed in response to virus infection and are potent inhibitors of STING palmitoylation and signaling.
- Author
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Hansen AL, Buchan GJ, Rühl M, Mukai K, Salvatore SR, Ogawa E, Andersen SD, Iversen MB, Thielke AL, Gunderstofte C, Motwani M, Møller CT, Jakobsen AS, Fitzgerald KA, Roos J, Lin R, Maier TJ, Goldbach-Mansky R, Miner CA, Qian W, Miner JJ, Rigby RE, Rehwinkel J, Jakobsen MR, Arai H, Taguchi T, Schopfer FJ, Olagnier D, and Holm CK
- Subjects
- Animals, Autoimmune Diseases of the Nervous System genetics, Autoimmune Diseases of the Nervous System metabolism, Autoimmune Diseases of the Nervous System pathology, Herpes Simplex genetics, Herpes Simplex pathology, Humans, Interferon Type I genetics, Interferon Type I metabolism, Lipoylation, Lupus Erythematosus, Systemic genetics, Lupus Erythematosus, Systemic metabolism, Lupus Erythematosus, Systemic pathology, Membrane Proteins genetics, Mice, Mice, Knockout, Nervous System Malformations genetics, Nervous System Malformations metabolism, Nervous System Malformations pathology, RAW 264.7 Cells, Fatty Acids metabolism, Herpes Simplex metabolism, Herpesvirus 2, Human metabolism, Membrane Proteins metabolism, Signal Transduction
- Abstract
The adaptor molecule stimulator of IFN genes (STING) is central to production of type I IFNs in response to infection with DNA viruses and to presence of host DNA in the cytosol. Excessive release of type I IFNs through STING-dependent mechanisms has emerged as a central driver of several interferonopathies, including systemic lupus erythematosus (SLE), Aicardi-Goutières syndrome (AGS), and stimulator of IFN genes-associated vasculopathy with onset in infancy (SAVI). The involvement of STING in these diseases points to an unmet need for the development of agents that inhibit STING signaling. Here, we report that endogenously formed nitro-fatty acids can covalently modify STING by nitro-alkylation. These nitro-alkylations inhibit STING palmitoylation, STING signaling, and subsequently, the release of type I IFN in both human and murine cells. Furthermore, treatment with nitro-fatty acids was sufficient to inhibit production of type I IFN in fibroblasts derived from SAVI patients with a gain-of-function mutation in STING. In conclusion, we have identified nitro-fatty acids as endogenously formed inhibitors of STING signaling and propose for these lipids to be considered in the treatment of STING-dependent inflammatory diseases., Competing Interests: Conflict of interest statement: F.J.S. declares financial interest in Complexa Inc., (Copyright © 2018 the Author(s). Published by PNAS.)
- Published
- 2018
- Full Text
- View/download PDF
48. Mitochondrial double-stranded RNA triggers antiviral signalling in humans.
- Author
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Dhir A, Dhir S, Borowski LS, Jimenez L, Teitell M, Rötig A, Crow YJ, Rice GI, Duffy D, Tamby C, Nojima T, Munnich A, Schiff M, de Almeida CR, Rehwinkel J, Dziembowski A, Szczesny RJ, and Proudfoot NJ
- Subjects
- Animals, DEAD-box RNA Helicases deficiency, DEAD-box RNA Helicases genetics, DEAD-box RNA Helicases metabolism, Endoribonucleases metabolism, Exoribonucleases deficiency, Exoribonucleases genetics, Exoribonucleases metabolism, Gene Expression Regulation immunology, HeLa Cells, Herpesvirus 1, Human genetics, Humans, Interferon Type I antagonists & inhibitors, Interferon Type I immunology, Interferon-Induced Helicase, IFIH1 metabolism, Mice, Mice, Inbred C57BL, Multienzyme Complexes metabolism, Mutation, Polyribonucleotide Nucleotidyltransferase metabolism, RNA Helicases metabolism, Single-Cell Analysis, bcl-2 Homologous Antagonist-Killer Protein metabolism, bcl-2-Associated X Protein metabolism, Herpesvirus 1, Human immunology, RNA, Double-Stranded immunology, RNA, Mitochondrial immunology
- Abstract
Mitochondria are descendants of endosymbiotic bacteria and retain essential prokaryotic features such as a compact circular genome. Consequently, in mammals, mitochondrial DNA is subjected to bidirectional transcription that generates overlapping transcripts, which are capable of forming long double-stranded RNA structures
1,2 . However, to our knowledge, mitochondrial double-stranded RNA has not been previously characterized in vivo. Here we describe the presence of a highly unstable native mitochondrial double-stranded RNA species at single-cell level and identify key roles for the degradosome components mitochondrial RNA helicase SUV3 and polynucleotide phosphorylase PNPase in restricting the levels of mitochondrial double-stranded RNA. Loss of either enzyme results in massive accumulation of mitochondrial double-stranded RNA that escapes into the cytoplasm in a PNPase-dependent manner. This process engages an MDA5-driven antiviral signalling pathway that triggers a type I interferon response. Consistent with these data, patients carrying hypomorphic mutations in the gene PNPT1, which encodes PNPase, display mitochondrial double-stranded RNA accumulation coupled with upregulation of interferon-stimulated genes and other markers of immune activation. The localization of PNPase to the mitochondrial inter-membrane space and matrix suggests that it has a dual role in preventing the formation and release of mitochondrial double-stranded RNA into the cytoplasm. This in turn prevents the activation of potent innate immune defence mechanisms that have evolved to protect vertebrates against microbial and viral attack.- Published
- 2018
- Full Text
- View/download PDF
49. Infection with a Brazilian isolate of Zika virus generates RIG-I stimulatory RNA and the viral NS5 protein blocks type I IFN induction and signaling.
- Author
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Hertzog J, Dias Junior AG, Rigby RE, Donald CL, Mayer A, Sezgin E, Song C, Jin B, Hublitz P, Eggeling C, Kohl A, and Rehwinkel J
- Subjects
- Active Transport, Cell Nucleus, Brazil, DEAD Box Protein 58 genetics, Down-Regulation, HEK293 Cells, Humans, Interferon Type I genetics, Phosphorylation, Receptors, Immunologic, Signal Transduction, Virus Replication, Zika Virus, Zika Virus Infection, DEAD Box Protein 58 immunology, Interferon Type I metabolism, RNA immunology, STAT1 Transcription Factor metabolism, STAT2 Transcription Factor metabolism, Viral Nonstructural Proteins metabolism
- Abstract
Zika virus (ZIKV) is a major public health concern in the Americas. We report that ZIKV infection and RNA extracted from ZIKV infected cells potently activated the induction of type I interferons (IFNs). This effect was fully dependent on the mitochondrial antiviral signaling protein (MAVS), implicating RIG-I-like receptors (RLRs) as upstream sensors of viral RNA. Indeed, RIG-I and the related RNA sensor MDA5 contributed to type I IFN induction in response to RNA from infected cells. We found that ZIKV NS5 from a recent Brazilian isolate blocked type I IFN induction downstream of RLRs and also inhibited type I IFN receptor (IFNAR) signaling. We defined the ZIKV NS5 nuclear localization signal and report that NS5 nuclear localization was not required for inhibition of signaling downstream of IFNAR. Mechanistically, NS5 blocked IFNAR signaling by both leading to reduced levels of STAT2 and by blocking phosphorylation of STAT1, two transcription factors activated by type I IFNs. Taken together, our observations suggest that ZIKV infection induces a type I IFN response via RLRs and that ZIKV interferes with this response by blocking signaling downstream of RLRs and IFNAR., (© 2018 The Authors. European Journal of Immunology published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2018
- Full Text
- View/download PDF
50. B Cells Producing Type I IFN Modulate Macrophage Polarization in Tuberculosis.
- Author
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Bénard A, Sakwa I, Schierloh P, Colom A, Mercier I, Tailleux L, Jouneau L, Boudinot P, Al-Saati T, Lang R, Rehwinkel J, Loxton AG, Kaufmann SHE, Anton-Leberre V, O'Garra A, Sasiain MDC, Gicquel B, Fillatreau S, Neyrolles O, and Hudrisier D
- Subjects
- Animals, Disease Models, Animal, Humans, Lung metabolism, Lung microbiology, Mice, Mice, Inbred C57BL, Mycobacterium tuberculosis, Signal Transduction, Spleen metabolism, Spleen microbiology, B-Lymphocytes metabolism, Interferon Type I metabolism, Macrophages metabolism, Tuberculosis metabolism
- Abstract
Rationale: In addition to their well-known function as antibody-producing cells, B lymphocytes can markedly influence the course of infectious or noninfectious diseases via antibody-independent mechanisms. In tuberculosis (TB), B cells accumulate in lungs, yet their functional contribution to the host response remains poorly understood., Objectives: To document the role of B cells in TB in an unbiased manner., Methods: We generated the transcriptome of B cells isolated from Mycobacterium tuberculosis (Mtb)-infected mice and validated the identified key pathways using in vitro and in vivo assays. The obtained data were substantiated using B cells from pleural effusion of patients with TB., Measurements and Main Results: B cells isolated from Mtb-infected mice displayed a STAT1 (signal transducer and activator of transcription 1)-centered signature, suggesting a role for IFNs in B-cell response to infection. B cells stimulated in vitro with Mtb produced type I IFN, via a mechanism involving the innate sensor STING (stimulator of interferon genes), and antagonized by MyD88 (myeloid differentiation primary response 88) signaling. In vivo, B cells expressed type I IFN in the lungs of Mtb-infected mice and, of clinical relevance, in pleural fluid from patients with TB. Type I IFN expression by B cells induced an altered polarization of macrophages toward a regulatory/antiinflammatory profile in vitro. In vivo, increased provision of type I IFN by B cells in a murine model of B cell-restricted Myd88 deficiency correlated with an enhanced accumulation of regulatory/antiinflammatory macrophages in Mtb-infected lungs., Conclusions: Type I IFN produced by Mtb-stimulated B cells favors macrophage polarization toward a regulatory/antiinflammatory phenotype during Mtb infection.
- Published
- 2018
- Full Text
- View/download PDF
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