bims-tunefa 2020-06-28 papers

bims-tunefa

Biomed News

on Tumor necrosis factor superfamily and post-translational modifications

Issue of 2020‒06‒28
twenty-six papers selected by
John Silke
Walter and Eliza Hall Institute of Medical Research

The deubiquitinase USP7 uses a distinct ubiquitin-like domain to deubiquitinate NF-κB subunits.
A Nimbolide-Based Kinase Degrader Preferentially Degrades Oncogenic BCR-ABL.
Loss of NFKB1 Results in Expression of Tumor Necrosis Factor and Activation of STAT1 to Promote Gastric Tumorigenesis in Mice.
Regulation of Interferon Induction by the Ubiquitin-Like Modifier FAT10.
Isoleucine 44 Hydrophobic Patch Controls Toxicity of Unanchored, Linear Ubiquitin Chains through NF-κB Signaling.
Regulating RIPK1: another way in which ULK1 contributes to survival.
The multiple roles of the IAP super-family in cancer.
Cross talk between intracellular pathogens and cell death.
The Immunoglobulin Superfamily Receptome Defines Cancer-Relevant Networks Associated with Clinical Outcome.
Identification and characterization of diverse OTU deubiquitinases in bacteria.
Defining the Threshold IL-2 Signal Required for Induction of Selective Treg Cell Responses Using Engineered IL-2 Muteins.
Ubiquitination of interleukin-1α is associated with increased pro-inflammatory polarization of murine macrophages deficient in the E3 ligase ITCH.
Loss of NF-kB1 and c-Rel accelerates oral carcinogenesis in mice.
Targeted deletion of interleukin-6 in a mouse model of chronic inflammation demonstrates opposing roles in aging: benefit and harm.
Jack of all trades inhibits inflammation (in sober people).
C1q/TNF-related Protein 1, a Multifunctional Adipokine: An Overview of Current Data.
TNF deficiency dysregulates inflammatory cytokine production, leading to lung pathology and death during respiratory poxvirus infection.
Evolution-inspired design of multicolored photoswitches from a single cyanobacteriochrome scaffold.
The multiple ways Wnt signaling contributes to acute leukemia pathogenesis.
ERK signalling: a master regulator of cell behaviour, life and fate.
Inflammasomes.
Whole-genome sequencing of patients with rare diseases in a national health system.
Comparing the efficacy and tolerability of biologic therapies in psoriasis: an updated network meta-analysis.
Interplay between RAGE and TLR4 Regulates HMGB1-Induced Inflammation by Promoting Cell Surface Expression of RAGE and TLR4.
SnapShot: Jak-STAT Signaling II.
SIX1 cooperates with RUNX1 and SMAD4 in cell fate commitment of Müllerian duct epithelium.

J Biol Chem. 2020 Jun 25. pii: jbc.RA120.014113. [Epub ahead of print]

The deubiquitinase USP7 uses a distinct ubiquitin-like domain to deubiquitinate NF-κB subunits.

Izaskun Mitxitorena, Domenico Somma, Jennifer P Mitchell, Matti Lepistö, Christian Tyrchan, Emma L Smith, Patrick A Kiely, Helen Walden, Karen Keeshan, Ruaidhrí J Carmody.

  The transcription factor NF-ĸB is a master regulator of the innate immune response and plays a central role in inflammatory diseases by mediating the expression of pro-inflammatory cytokines. Ubiquitination-triggered proteasomal degradation of DNA-bound NF-ĸB strongly limits the expression of its target genes. Conversely, the deubiquitinase ubiquitin-specific peptidase 7 (USP7) opposes the activities of E3 ligases, stabilizes DNA-bound NF-ĸB, and thereby promotes NF-ĸB-mediated transcription. Using gene expression and synthetic peptide arrays on membrane support (SPOT) synthesis of peptides and overlay analyses, we found here that inhibiting USP7 increases NF-ĸB ubiquitination and degradation, prevents Toll-like receptor-induced pro-inflammatory cytokine expression, and represents an effective strategy for controlling inflammation. However, the broad regulatory roles of USP7 in cell death pathways, chromatin, and DNA damage responses limits the use of catalytic inhibitors of USP7 as anti-inflammatory agents. To this end, we identified an NF-ĸB-binding site in USP7, ubiquitin-like domain 2, that selectively mediates interactions of USP7 with NF-ĸB subunits, but is dispensable for interactions with other proteins. Moreover, we found that the amino acids 757LDEL760 in USP7 critically contribute to the interaction with the p65 subunit of NF-ĸB. Our findings support the notion that USP7 activity could be potentially targeted in a substrate-selective manner through the development of non-catalytic inhibitors of this deubiquitinase to abrogate NF-ĸB activity.

Keywords:  NF-kappa B (NF-KB); deubiquitinase; deubiquitylation (deubiquitination); gene regulation; inflammation; innate immunity; protein-protein interaction; toll-like receptor (TLR); transcription factor; ubiquitin like domain; ubiquitin specific peptidase 7 (USP7); ubiquitylation (ubiquitination)

DOI:  https://doi.org/10.1074/jbc.RA120.014113
ACS Chem Biol. 2020 Jun 25.

A Nimbolide-Based Kinase Degrader Preferentially Degrades Oncogenic BCR-ABL.

Bingqi Tong, Jessica N Spradlin, Luiz F T Novaes, Erika Zhang, Xirui Hu, Malte Moeller, Scott M Brittain, Lynn M McGregor, Jeffrey M McKenna, John A Tallarico, Markus Schirle, Thomas J Maimone, Daniel K Nomura.

Targeted protein degradation (TPD) and proteolysis-targeting chimeras (PROTACs) have arisen as powerful therapeutic modalities for degrading specific proteins in a proteasome-dependent manner. However, a major limitation of TPD is the lack of E3 ligase recruiters. Recently, we discovered the natural product nimbolide as a covalent recruiter for the E3 ligase RNF114. Here, we show the broader utility of nimbolide as an E3 ligase recruiter for TPD applications. We demonstrate that a PROTAC linking nimbolide to the kinase and BCR-ABL fusion oncogene inhibitor dasatinib, BT1, selectively degrades BCR-ABL over c-ABL in leukemia cancer cells, compared to previously reported cereblon or VHL-recruiting BCR-ABL degraders that show opposite selectivity or, in some cases, inactivity. Thus, we further establish nimbolide as an additional general E3 ligase recruiter for PROTACs, and we demonstrate the importance of expanding upon the arsenal of E3 ligase recruiters, as such molecules confer differing selectivity for the degradation of neo-substrate proteins.

DOI: https://doi.org/10.1021/acschembio.0c00348
Gastroenterology. 2020 Jun 19. pii: S0016-5085(20)34840-X. [Epub ahead of print]

Loss of NFKB1 Results in Expression of Tumor Necrosis Factor and Activation of STAT1 to Promote Gastric Tumorigenesis in Mice.

Jun T Low, Michael Christie, Matthias Ernst, Laure Dumoutier, Adele Preaudet, Yanhong Ni, Michael D W Griffin, Lisa A Mielke, Andreas Strasser, Tracy L Putoczki, Lorraine A O'Reilly.

  BACKGROUND & AIMS: Activity of NFkB transcription factors and signaling via STAT are frequently altered in gastric cancer cells. Mice lacking NFKB1 (Nfkb1-/- mice) develop invasive gastric cancer and their gastric tissues have increased levels of cytokines, such as interleukin (IL)6, IL22, IL11, and tumor necrosis factor (TNF), as well as increased activation of signal transducer and activator of transcription 1 (STAT1). We investigated whether these cytokines were required for STAT1 activation in gastric tissues of mice and critical for gastric tumorigenesis.METHODS: We crossed Nfkb1-/- mice with Il6-/-, Il22-/-, Il11Rα-/- and Tnf-/- mice. Stomach tissues from compound mutant mice were analyzed by histology, immunoblotting and RNA sequencing. Lymphoid, myeloid and epithelial cells were isolated from stomachs and the levels of cytokines were determined by flow cytometric analysis.
RESULTS: Nfkb1-/- mice developed gastritis, oxyntic atrophy, gastric dysplasia and invasive tumors, whereas Nfkb1-/-Stat1-/- mice did not, even when followed for as long as 2 years. The levels of Il6, Il11, and Il22 and Tnf mRNA were increased in the body and antrum of the stomachs from Nfkb1-/- mice, from 6 months of age. However, Nfkb1-/-Il6-/-, Nfkb1-/-Il22-/- and Nfkb1-/-Il11Rα-/- mice still developed gastric tumors, although the absence of IL11 receptor (IL11R) significantly reduced development of invasive gastric tumors. Stomachs from Nfkb1-/-Tnf-/- mice exhibited significantly less gastritis and oxyntic atrophy and fewer tumors than Nfkb1-/- mice. This correlated with reduced activation of STAT1 and STAT3 and fewer numbers of T cells and B cells infiltrating the gastric body. Loss of STAT1 significantly reduced expression of PD-L1 on epithelial and myeloid (CD11b+) cells in the gastric mucosa of Nfkb1-/- mice, indeed to the levels observed on the corresponding cells from wild-type mice.
CONCLUSIONS: In studies of gastric tumor development in knockout mice, we found that loss of NFKB1 causes increased expression of TNF in the stomach and thereby drives activation of STAT1, resulting in an inflammatory immune response and the development of gastric cancer. IL11R appears to be required for the progression of gastric tumors to the invasive stage. These findings suggest that inhibitors of TNF, and possibly also inhibitors of IL11/IL11Rα, might be useful in the treatment of gastric cancer.

Keywords:  inflammation; inhibitory signal; monocyte; programmed cell death 1 ligand 1

DOI:  https://doi.org/10.1053/j.gastro.2020.06.039
Biomolecules. 2020 Jun 23. pii: E951. [Epub ahead of print]10(6):

Regulation of Interferon Induction by the Ubiquitin-Like Modifier FAT10.

Mei Min Mah, Nicola Roverato, Marcus Groettrup.

  The revelation that the human major histocompatibility complex (MHC) class I locus encodes a ubiquitin-like protein designated HLA-F adjacent transcript 10 (FAT10) or ubiquitin D (UBD) has attracted increasing attention to the function of this protein. Interestingly, the pro-inflammatory cytokines interferon (IFN)-γ and tumor necrosis factor (TNF) α synergize to strongly induce FAT10 expression, thereby suggesting a role of FAT10 in the immune response. Recent reports that FAT10 downregulates type I interferon production while it upregulates IFN-γ pose mechanistic questions on how FAT10 differentially regulates interferon induction. Several covalent and non-covalent binding partners of FAT10 involved in signal transduction pathways leading to IFN synthesis have been identified. After introducing FAT10, we review here recent insights into how FAT10 affects proteins in the interferon pathways, like the virus-responsive pattern recognition receptor RIG-I, the ubiquitin ligase ZNF598, and the deubiquitylating enzyme OTUB1. Moreover, we outline the consequences of FAT10 deficiency on interferon synthesis and viral expansion in mice and human cells. We discuss the need for covalent isopeptide linkage of FAT10 to the involved target proteins and the concomitant targeting for proteasomal degradation. After years of investigating the elusive biological functions of this fascinating ubiquitin-like modifier, we review the emerging evidence for a novel role of FAT10 in interferon regulation.

Keywords:  FAT10; RIG-I; influenza virus; interferon; signal transduction; ubiquitin

DOI:  https://doi.org/10.3390/biom10060951
Cells. 2020 Jun 22. pii: E1519. [Epub ahead of print]9(6):

Isoleucine 44 Hydrophobic Patch Controls Toxicity of Unanchored, Linear Ubiquitin Chains through NF-κB Signaling.

Jessica R Blount, Kozeta Libohova, Gustavo M Silva, Sokol V Todi.

  Ubiquitination is a post-translational modification that regulates cellular processes by altering the interactions of proteins to which ubiquitin, a small protein adduct, is conjugated. Ubiquitination yields various products, including mono- and poly-ubiquitinated substrates, as well as unanchored poly-ubiquitin chains whose accumulation is considered toxic. We previously showed that transgenic, unanchored poly-ubiquitin is not problematic in Drosophila melanogaster. In the fruit fly, free chains exist in various lengths and topologies and are degraded by the proteasome; they are also conjugated onto other proteins as one unit, eliminating them from the free ubiquitin chain pool. Here, to further explore the notion of unanchored chain toxicity, we examined when free poly-ubiquitin might become problematic. We found that unanchored chains can be highly toxic if they resemble linear poly-ubiquitin that cannot be modified into other topologies. These species upregulate NF-κB signaling, and modulation of the levels of NF-κB components reduces toxicity. In additional studies, we show that toxicity from untethered, linear chains is regulated by isoleucine 44, which anchors a key interaction site for ubiquitin. We conclude that free ubiquitin chains can be toxic, but only in uncommon circumstances, such as when the ability of cells to modify and regulate them is markedly restricted.

Keywords:  Drosophila melanogaster; NF-κB signaling; innate immunity; ubiquitination; unanchored poly-ubiquitin

DOI:  https://doi.org/10.3390/cells9061519
Autophagy. 2020 Jun 24. 1-3

Regulating RIPK1: another way in which ULK1 contributes to survival.

Wenxian Wu, Björn Stork.

  The mammalian ULK1 is the central initiating kinase of bulk and selective macroautophagy/autophagy processes. In the past, both autophagy-relevant and non-autophagy-relevant substrates of this Ser/Thr kinase have been reported. Here, we describe our recent finding that ULK1 also regulates TNF signaling pathways. We find that inhibition of autophagy or specifically ULK1 increases TNF-induced cell death. This autophagy-independent pro-survival function of ULK1 is mediated via the phosphorylation of RIPK1 at Ser357. RIPK1 is the central mediator of pro-inflammatory or pro-death signaling pathways induced by TNF, and ULK1-dependent phosphorylation regulates RIPK1 activation and distribution to different intracellular signaling complexes. Our results indicate that ULK1 exerts a cyto-protective function not only by initiating autophagy, but also by controlling RIPK1-mediated cell death.

Keywords:  Autophagy; RIPK1; TNF; ULK1; necroptosis; necrosome

DOI:  https://doi.org/10.1080/15548627.2020.1783110
Pharmacol Ther. 2020 Jun 22. pii: S0163-7258(20)30140-6. [Epub ahead of print] 107610

The multiple roles of the IAP super-family in cancer.

Swati Kumar, Ciaran Fairmichael, Daniel B Longley, Richard C Turkington.

  The Inhibitor of Apoptosis proteins (IAPs) are a family of proteins that are mainly known for their anti-apoptotic activity and ability to directly bind and inhibit caspases. Recent research has however revealed that they have extensive roles in governing numerous other cellular processes. IAPs are known to modulate ubiquitin (Ub)-dependent signaling pathways through their E3 ligase activity and influence activation of nuclear factor κB (NFκB). In this review, we discuss the involvement of IAPs in individual hallmarks of cancer and the current status of therapies targeting these critical proteins.

Keywords:  Cancer therapy; Hallmarks of cancer; Inhibitors of apoptosis; Molecular targets

DOI:  https://doi.org/10.1016/j.pharmthera.2020.107610
Immunol Rev. 2020 Jun 22.

Cross talk between intracellular pathogens and cell death.

Benjamin Demarco, Kaiwen W Chen, Petr Broz.

  Infections with bacterial pathogens often results in the initiation of programmed cell death as part of the host innate immune defense, or as a bacterial virulence strategy. Induction of host cell death is controlled by an elaborate network of innate immune and cell death signaling pathways and manifests in different morphologically and functionally distinct forms of death, such as apoptosis, necroptosis, NETosis and pyroptosis. The mechanism by which host cell death restricts bacterial replication is highly cell-type and context depended, but its physiological importance is highlighted the diversity of strategies bacterial pathogens use to avoid induction of cell death or to block cell death signaling pathways. In this review, we discuss the latest insights into how bacterial pathogens elicit and manipulate cell death signaling, how different forms of cell death kill or restrict bacteria and how cell death and innate immune pathway cross talk to guard against pathogen-induced inhibition of host cell death.

Keywords:  apoptosis; infection; innate immunity; necroptosis; programmed cell death; pyroptosis

DOI:  https://doi.org/10.1111/imr.12892
Cell. 2020 Jun 23. pii: S0092-8674(20)30694-2. [Epub ahead of print]

The Immunoglobulin Superfamily Receptome Defines Cancer-Relevant Networks Associated with Clinical Outcome.

Erik Verschueren, Bushra Husain, Kobe Yuen, Yi Sun, Sairupa Paduchuri, Yasin Senbabaoglu, Isabelle Lehoux, Tia A Arena, Blair Wilson, Steve Lianoglou, Corey Bakalarski, Yvonne Franke, Pamela Chan, Athena W Wong, Lino C Gonzalez, Sanjeev Mariathasan, Shannon J Turley, Jennie R Lill, Nadia Martinez-Martin.

  Cell surface receptors and their interactions play a central role in physiological and pathological signaling. Despite its clinical relevance, the immunoglobulin superfamily (IgSF) remains uncharacterized and underrepresented in databases. Here, we present a systematic extracellular protein map, the IgSF interactome. Using a high-throughput technology to interrogate most single transmembrane receptors for binding to 445 IgSF proteins, we identify over 500 interactions, 82% previously undocumented, and confirm more than 60 receptor-ligand pairs using orthogonal assays. Our study reveals a map of cell-type-specific interactions and the landscape of dysregulated receptor-ligand crosstalk in cancer, including selective loss of function for tumor-associated mutations. Furthermore, investigation of the IgSF interactome in a large cohort of cancer patients identifies interacting protein signatures associated with clinical outcome. The IgSF interactome represents an important resource to fuel biological discoveries and a framework for understanding the functional organization of the surfaceome during homeostasis and disease, ultimately informing therapeutic development.

Keywords:  cancer networks; cell surface; extracellular interactome; immunoglobulin superfamily; receptor-ligand interactions

DOI:  https://doi.org/10.1016/j.cell.2020.06.007
EMBO J. 2020 Jun 22. e105127

Identification and characterization of diverse OTU deubiquitinases in bacteria.

Alexander F Schubert, Justine V Nguyen, Tyler G Franklin, Paul P Geurink, Cameron G Roberts, Daniel J Sanderson, Lauren N Miller, Huib Ovaa, Kay Hofmann, Jonathan N Pruneda, David Komander.

  Manipulation of host ubiquitin signaling is becoming an increasingly apparent evolutionary strategy among bacterial and viral pathogens. By removing host ubiquitin signals, for example, invading pathogens can inactivate immune response pathways and evade detection. The ovarian tumor (OTU) family of deubiquitinases regulates diverse ubiquitin signals in humans. Viral pathogens have also extensively co-opted the OTU fold to subvert host signaling, but the extent to which bacteria utilize the OTU fold was unknown. We have predicted and validated a set of OTU deubiquitinases encoded by several classes of pathogenic bacteria. Biochemical assays highlight the ubiquitin and polyubiquitin linkage specificities of these bacterial deubiquitinases. By determining the ubiquitin-bound structures of two examples, we demonstrate the novel strategies that have evolved to both thread an OTU fold and recognize a ubiquitin substrate. With these new examples, we perform the first cross-kingdom structural analysis of the OTU fold that highlights commonalities among distantly related OTU deubiquitinases.

Keywords:  bacterial effector; deubiquitinase; pathogen; protein structure; ubiquitin

DOI:  https://doi.org/10.15252/embj.2020105127
Front Immunol. 2020 ;11 1106

Defining the Threshold IL-2 Signal Required for Induction of Selective Treg Cell Responses Using Engineered IL-2 Muteins.

Aazam Ghelani, Darren Bates, Kip Conner, Min-Zu Wu, Jiamiao Lu, Yi-Ling Hu, Chi-Ming Li, Ashutosh Chaudhry, Sue J Sohn.

  Among all T and NK cell subsets, regulatory T (Treg) cells typically respond to the lowest concentrations of IL-2 due to elevated surface expression of the IL-2R alpha chain (IL2RA; CD25) and the high affinity IL-2 receptor (IL-2R) complex. This enhanced sensitivity forms the basis for low-dose (LD) IL-2 therapy for the treatment of inflammatory diseases, where efficacy correlates with increased Treg cell number and expression of functional markers. Despite strong preclinical support for this approach, moderate and variable clinical efficacy has raised concerns that adequate Treg selectivity still cannot be achieved with LD IL-2, and/or that doses are too low to stimulate effective Treg-mediated suppression within tissues. This has prompted development of IL-2 variants with greater Treg selectivity, achieved through attenuated affinity for the signaling chains of the IL-2R complex (IL2RB or CD122 and IL2RG or CD132) and, consequently, greater reliance on high CD25 levels for full receptor binding and signaling. While certain IL-2 variants have advanced to the clinic, it remains unknown if the full range of IL-2R signaling potency and Treg-selectivity observed with low concentrations of wildtype IL-2 can be sufficiently recapitulated with attenuated IL-2 muteins at high concentrations. Using a panel of engineered IL-2 muteins, we investigated how a range of IL-2R signaling intensity, benchmarked by the degree of STAT5 phosphorylation, relates to biologically relevant Treg cell responses such as proliferation, lineage and phenotypic marker expression, and suppressor function. Our results demonstrate that a surprisingly wide dynamic range of IL-2R signaling intensity leads to productive biological responses in Treg cells, with negligible STAT5 phosphorylation associating with nearly complete downstream effects such as Treg proliferation and suppressor activity. Furthermore, we show with both in vitro and humanized mouse in vivo systems that different biological responses in Treg cells require different minimal IL-2R signaling thresholds. Our findings suggest that more than minimal IL-2R signaling, beyond that capable of driving Treg cell proliferation, may be required to fully enhance Treg cell stability and suppressor function in vivo.

Keywords:  IL-2; Treg; autoimmunity; inflammation; mutein; regulatory T; tolerance

DOI:  https://doi.org/10.3389/fimmu.2020.01106
J Biol Chem. 2020 Jun 25. pii: jbc.RA120.014298. [Epub ahead of print]

Ubiquitination of interleukin-1α is associated with increased pro-inflammatory polarization of murine macrophages deficient in the E3 ligase ITCH.

Xi Lin, Hengwei Zhang, Brendan F Boyce, Lianping Xing.

  Macrophages play critical roles in homeostasis and inflammation. Macrophage polarization to either a pro-inflammatory or anti-inflammatory status is controlled by activating inflammatory signaling pathways. Ubiquitination is a post-translational modification that regulates these inflammatory signaling pathways. However, the influence of protein ubiquitination on macrophage polarization has not been well studied. We hypothesized that the ubiquitination status of key proteins in inflammatory pathways contributes to macrophage polarization, which is regulated by itchy E3 ubiquitin ligase (ITCH), a negative regulator of inflammation. Using ubiquitin proteomics, we found that ubiquitination profiles are different among polarized murine macrophage subsets. Interestingly, interleukin-1α (IL-1α), an important proinflammatory mediator, was specifically ubiquitinated in lipopolysaccharide-induced proinflammatory macrophages, which was enhanced in ITCH-deficient macrophages. The ITCH-deficient macrophages had increased levels of the mature form of IL-1α and exhibited proinflammatory polarization, and reduced deubiquitination of IL-1α protein. Finally, IL-1α neutralization attenuated pro-inflammatory polarization of the ITCH-deficient macrophages. In conclusion, ubiquitination of IL-1α is associated with increased pro-inflammatory polarization of macrophages deficient in the E3 ligase ITCH.

Keywords:  Interleukin-1α; Itch; deubiquitylation (deubiquitination); inflammation; interleukin 1 (IL-1); macrophage; macrophage polarization; ubiquitylation (ubiquitination)

DOI:  https://doi.org/10.1074/jbc.RA120.014298
Oral Dis. 2020 Jun 22.

Loss of NF-kB1 and c-Rel accelerates oral carcinogenesis in mice.

Yanhong Ni, Tami Yap, Natasha Silke, John Silke, Michael McCullough, Antonio Celentano, Lorraine A O'Reilly.



Keywords:  NF-kB1; c-Rel; carcinogenesis; inflammation; mouth neoplasms; oral

DOI:  https://doi.org/10.1111/odi.13508
J Gerontol A Biol Sci Med Sci. 2020 Jun 26. pii: glaa156. [Epub ahead of print]

Targeted deletion of interleukin-6 in a mouse model of chronic inflammation demonstrates opposing roles in aging: benefit and harm.

Lina Ma, Lolita S Nidadavolu, Huanle Yang, Jackie Langdon, Reyhan Westbrook, Benjamin M W Tsui, Taek-Soo Lee, Jared Hinson, Shizhang Ling, Ruth Marx-Rattner, Yuqiong Wu, Tu Nguyen, Jessica Tan, Mohammed Khadeer, Ruin Moaddel, Anne Le, Jeremy D Walston, Peter M Abadir.

  Chronic inflammation (CI) in older adults is associated with reduced healthspan and lifespan. Interleukin-6 (IL-6) is one CI marker that is strongly associated with adverse health outcomes and mortality in aging. We have previously characterized a mouse model of frailty and chronic inflammatory pathway activation (IL-10tm/tm, IL-10 KO) that demonstrates the upregulation of numerous proinflammatory cytokines, including IL-6. We sought to identify a more specific role for IL-6 within the context of CI and aging and developed a mouse with targeted deletion of both IL-10 and IL-6 (IL-10tm/tm/IL-6tm/tm, DKO). Phenotypic characteristics, cytokine measurements, cardiac myocardial oxygen consumption, physical function, and survival were measured in DKO mice and compared to age- and gender-matched IL-10 KO and wild-type mice. Our findings demonstrate that selective knockdown of IL-6 in a frail mouse with CI resulted in the reversal of some of the CI-associated changes. We observed increased protective mitochondrial-associated lipid metabolites, decreased cardiac oxaloacetic acid, improved myocardial oxidative metabolism, and better short-term functional performance in DKO mice. However, the DKO mice also demonstrated higher mortality. This work shows the pleiotropic effects of IL-6 on aging and frailty.

Keywords:  Interleukin-6; interleukin-10; knockout; lysophosphatidylcholine; mitochondria

DOI:  https://doi.org/10.1093/gerona/glaa156
Nat Immunol. 2020 Jul;21(7): 718-719

Jack of all trades inhibits inflammation (in sober people).

Florian I Schmidt, Eicke Latz.

DOI: https://doi.org/10.1038/s41590-020-0693-6
Am J Med Sci. 2020 May 28. pii: S0002-9629(20)30225-1. [Epub ahead of print]

C1q/TNF-related Protein 1, a Multifunctional Adipokine: An Overview of Current Data.

Joanna Dorota Janowska.

  The present review aimed to present the research highlights on C1q/TNF-related protein 1 (CTRP1), a member of the recently discovered family of highly conserved adiponectin paralog proteins, C1q tumor necrosis factor-related proteins. CTRP1 plays an important role in regulating body energy homeostasis and sensitivity to insulin. Studies on animal models have shown that it lowers the concentration of glucose. Elevated concentrations of CTRP1 reduce weight gain and diet-induced insulin resistance. CTRP1 limits the extent of ischemia-reperfusion injury in acute myocardial infarction. It inhibits platelet aggregation by blocking von Willebrand factor binding to collagen. In patients with chronic kidney disease, an increase in CTRP1 levels is associated with a lesser degree of disease progression. CTRP1 stimulates aldosterone synthesis in the adrenal cortex by affecting aldosterone synthase expression. In dehydration, an increase in CTRP1 concentration helps to maintain normotension. It participates in processes related to the proliferation and maturation of chondrocytes. It also promotes atherosclerosis, and a surge in its concentration is correlated with a higher cardiovascular risk in patients with coronary atherosclerosis. In vascular smooth muscle cells, it induces the expression of proinflammatory cytokines. An increase in CTRP1 levels is correlated with the progression of the neoplastic process in patients with glioblastoma.

Keywords:  C1qTNF-related protein 1; Chronic kidney disease; Coronary artery disease; Glucose and lipid metabolism; Inflammation

DOI:  https://doi.org/10.1016/j.amjms.2020.05.036
Proc Natl Acad Sci U S A. 2020 Jun 22. pii: 202004615. [Epub ahead of print]

TNF deficiency dysregulates inflammatory cytokine production, leading to lung pathology and death during respiratory poxvirus infection.

Ma Junaliah Tuazon Kels, Esther Ng, Zahrah Al Rumaih, Pratikshya Pandey, Sigrid R Ruuls, Heinrich Korner, Timothy P Newsome, Geeta Chaudhri, Gunasegaran Karupiah.

  Excessive tumor necrosis factor (TNF) is known to cause significant pathology. Paradoxically, deficiency in TNF (TNF-/-) also caused substantial pathology during respiratory ectromelia virus (ECTV) infection, a surrogate model for smallpox. TNF-/- mice succumbed to fulminant disease whereas wild-type mice, and those engineered to express only transmembrane TNF (mTNF), fully recovered. TNF deficiency did not affect viral load or leukocyte recruitment but caused severe lung pathology and excessive production of the cytokines interleukin (IL)-6, IL-10, transforming growth factor beta (TGF-β), and interferon gamma (IFN-γ). Short-term blockade of these cytokines significantly reduced lung pathology in TNF-/- mice concomitant with induction of protein inhibitor of activated STAT3 (PIAS3) and/or suppressor of cytokine signaling 3 (SOCS3), factors that inhibit STAT3 activation. Consequently, inhibition of STAT3 activation with an inhibitor reduced lung pathology. Long-term neutralization of IL-6 or TGF-β protected TNF-/- mice from an otherwise lethal infection. Thus, mTNF alone is necessary and sufficient to regulate lung inflammation but it has no direct antiviral activity against ECTV. The data indicate that targeting specific cytokines or cytokine-signaling pathways to reduce or ameliorate lung inflammation during respiratory viral infections is possible but that the timing and duration of the interventive measure are critical.

Keywords:  STAT3 dysregulation; dysregulated cytokine response; lung inflammation and pathology; respiratory poxvirus infection; tumor necrosis factor deficiency

DOI:  https://doi.org/10.1073/pnas.2004615117
Proc Natl Acad Sci U S A. 2020 Jun 22. pii: 202004273. [Epub ahead of print]

Evolution-inspired design of multicolored photoswitches from a single cyanobacteriochrome scaffold.

Keiji Fushimi, Masumi Hasegawa, Takeru Ito, Nathan C Rockwell, Gen Enomoto, Ni-Ni -Win, J Clark Lagarias, Masahiko Ikeuchi, Rei Narikawa.

  Cyanobacteriochromes (CBCRs) are small, bistable linear tetrapyrrole (bilin)-binding light sensors which are typically found as modular components in multidomain cyanobacterial signaling proteins. The CBCR family has been categorized into many lineages that roughly correlate with their spectral diversity, but CBCRs possessing a conserved DXCF motif are found in multiple lineages. DXCF CBCRs typically possess two conserved Cys residues: a first Cys that remains ligated to the bilin chromophore and a second Cys found in the DXCF motif. The second Cys often forms a second thioether linkage, providing a mechanism to sense blue and violet light. DXCF CBCRs have been described with blue/green, blue/orange, blue/teal, and green/teal photocycles, and the molecular basis for some of this spectral diversity has been well established. We here characterize AM1_1499g1, an atypical DXCF CBCR that lacks the second cysteine residue and exhibits an orange/green photocycle. Based on prior studies of CBCR spectral tuning, we have successfully engineered seven AM1_1499g1 variants that exhibit robust yellow/teal, green/teal, blue/teal, orange/yellow, yellow/green, green/green, and blue/green photocycles. The remarkable spectral diversity generated by modification of a single CBCR provides a good template for multiplexing synthetic photobiology systems within the same cellular context, thereby bypassing the time-consuming empirical optimization process needed for multiple probes with different protein scaffolds.

Keywords:  circular dichroism; optogenetics; phytochrome

DOI:  https://doi.org/10.1073/pnas.2004273117
J Leukoc Biol. 2020 Jun 23.

The multiple ways Wnt signaling contributes to acute leukemia pathogenesis.

Sheila C Soares-Lima, Maria S Pombo-de-Oliveira, Flávia R G Carneiro.

  WNT proteins constitute a very conserved family of secreted glycoproteins that act as short-range ligands for signaling with critical roles in hematopoiesis, embryonic development, and tissue homeostasis. These proteins transduce signals via the canonical pathway, which is β-catenin-mediated and better-characterized, or via more diverse noncanonical pathways that are β-catenin independent and comprise the planar cell polarity (PCP) pathway and the WNT/Ca++ pathways. Several proteins regulate Wnt signaling through a variety of sophisticated mechanisms. Disorders within the pathway can contribute to various human diseases, and the dysregulation of Wnt pathways by different molecular mechanisms is implicated in the pathogenesis of many types of cancer, including the hematological malignancies. The types of leukemia differ considerably and can be subdivided into chronic, myeloid or lymphocytic, and acute, myeloid or lymphocytic, leukemia, according to the differentiation stage of the predominant cells, the progenitor lineage, the diagnostic age strata, and the specific molecular drivers behind their development. Here, we review the role of Wnt signaling in normal hematopoiesis and discuss in detail the multiple ways canonical Wnt signaling can be dysregulated in acute leukemia, including alterations in gene expression and protein levels, epigenetic regulation, and mutations. Furthermore, we highlight the different impacts of these alterations, considering the distinct forms of the disease, and the therapeutic potential of targeting Wnt signaling.

Keywords:  AML; BCP-ALL; T-ALL; cancer; β-catenin

DOI:  https://doi.org/10.1002/JLB.2MR0420-707R
Nat Rev Mol Cell Biol. 2020 Jun 23.

ERK signalling: a master regulator of cell behaviour, life and fate.

Hugo Lavoie, Jessica Gagnon, Marc Therrien.

The proteins extracellular signal-regulated kinase 1 (ERK1) and ERK2 are the downstream components of a phosphorelay pathway that conveys growth and mitogenic signals largely channelled by the small RAS GTPases. By phosphorylating widely diverse substrates, ERK proteins govern a variety of evolutionarily conserved cellular processes in metazoans, the dysregulation of which contributes to the cause of distinct human diseases. The mechanisms underlying the regulation of ERK1 and ERK2, their mode of action and their impact on the development and homeostasis of various organisms have been the focus of much attention for nearly three decades. In this Review, we discuss the current understanding of this important class of kinases. We begin with a brief overview of the structure, regulation, substrate recognition and subcellular localization of ERK1 and ERK2. We then systematically discuss how ERK signalling regulates six fundamental cellular processes in response to extracellular cues. These processes are cell proliferation, cell survival, cell growth, cell metabolism, cell migration and cell differentiation.

DOI: https://doi.org/10.1038/s41580-020-0255-7
Curr Biol. 2020 Jun 22. pii: S0960-9822(20)30581-9. [Epub ahead of print]30(12): R689-R694

Inflammasomes.

Natália Ketelut-Carneiro, Katherine A Fitzgerald.

Ketelut-Carneiro and Fitzgerald discuss the basic concepts about inflammasome composition, assembly and activation, effector functions and role in infection and inflammatory disease.

DOI: https://doi.org/10.1016/j.cub.2020.04.065
Nature. 2020 Jun 24.

Whole-genome sequencing of patients with rare diseases in a national health system.

Ernest Turro, William J Astle, Karyn Megy, Stefan Gräf, Daniel Greene, Olga Shamardina, Hana Lango Allen, Alba Sanchis-Juan, Mattia Frontini, Chantal Thys, Jonathan Stephens, Rutendo Mapeta, Oliver S Burren, Kate Downes, Matthias Haimel, Salih Tuna, Sri V V Deevi, Timothy J Aitman, David L Bennett, Paul Calleja, Keren Carss, Mark J Caulfield, Patrick F Chinnery, Peter H Dixon, Daniel P Gale, Roger James, Ania Koziell, Michael A Laffan, Adam P Levine, Eamonn R Maher, Hugh S Markus, Joannella Morales, Nicholas W Morrell, Andrew D Mumford, Elizabeth Ormondroyd, Stuart Rankin, Augusto Rendon, Sylvia Richardson, Irene Roberts, Noemi B A Roy, Moin A Saleem, Kenneth G C Smith, Hannah Stark, Rhea Y Y Tan, Andreas C Themistocleous, Adrian J Thrasher, Hugh Watkins, Andrew R Webster, Martin R Wilkins, Catherine Williamson, James Whitworth, Sean Humphray, David R Bentley, , Nathalie Kingston, Neil Walker, John R Bradley, Sofie Ashford, Christopher J Penkett, Kathleen Freson, Kathleen E Stirrups, F Lucy Raymond, Willem H Ouwehand.

Most patients with rare diseases do not receive a molecular diagnosis and the aetiological variants and causative genes for more than half such disorders remain to be discovered1. Here we used whole-genome sequencing (WGS) in a national health system to streamline diagnosis and to discover unknown aetiological variants in the coding and non-coding regions of the genome. We generated WGS data for 13,037 participants, of whom 9,802 had a rare disease, and provided a genetic diagnosis to 1,138 of the 7,065 extensively phenotyped participants. We identified 95 Mendelian associations between genes and rare diseases, of which 11 have been discovered since 2015 and at least 79 are confirmed to be aetiological. By generating WGS data of UK Biobank participants2, we found that rare alleles can explain the presence of some individuals in the tails of a quantitative trait for red blood cells. Finally, we identified four novel non-coding variants that cause disease through the disruption of transcription of ARPC1B, GATA1, LRBA and MPL. Our study demonstrates a synergy by using WGS for diagnosis and aetiological discovery in routine healthcare.

DOI: https://doi.org/10.1038/s41586-020-2434-2
Br J Dermatol. 2020 Jun 20.

Comparing the efficacy and tolerability of biologic therapies in psoriasis: an updated network meta-analysis.

S K Mahil, M C Ezejimofor, L S Exton, L Manounah, A D Burden, L C Coates, M de Brito, A McGuire, R Murphy, C M Owen, R Parslew, R T Woolf, Z Z N Yiu, O A Uthman, M F Mohd Mustapa, C H Smith.

BACKGROUND/OBJECTIVES: The rapid expansion of psoriasis biologics has led to an urgent need to understand their relative efficacy and tolerability to better inform treatment decisions and specifically, to inform guideline development.METHODS: We searched MEDLINE, PubMed, EMBASE and Cochrane databases for randomized controlled trials (RCTs),published up to 7th September 2018, of 11 licensed, NICE-approved biologics targeting TNF(adalimumab, etanercept, infliximab, certolizumab pegol), IL-12/IL-23p40 (ustekinumab), IL-17A (secukinumab, ixekizumab), IL-17RA (brodalumab) and IL-23p19 (guselkumab, tildrakizumab, risankizumab). Data were extracted and synthesised using RevMan 5.3. A frequentist network meta-analysis, conducted using Stata13 (StataCorp), ascertained direct/indirect evidence comparing biologics with one another, methotrexate, or placebo. This was combined with hierarchical cluster analyses to consider efficacy (PASI90 or PGA0/1; PASI75; DLQI improvement) and tolerability (drug withdrawal due to adverse events) outcomes at 10-16 weeks, followed by study quality, heterogeneity, and inconsistency assessments.
RESULTS: We identified 62 RCTs presenting data on direct comparisons (n=31,899 participants). All biologics were efficacious compared with placebo or methotrexate at 10-16 weeks. Hierarchical cluster analyses revealed that adalimumab, brodalumab, certolizumab pegol, guselkumab, risankizumab, secukinumab, tildrakizumab and ustekinumab were comparable with respect to high short-term efficacy and tolerability. Infliximab and ixekizumab clustered together, with high short-term efficacy but relatively lower tolerability compared with other agents, although the number of drug withdrawal events across the network was low, so these findings should be treated with caution.
CONCLUSIONS: Using our methodology, we find most biologics cluster together with respect to short-term efficacy and tolerability, and we do not identify any single agent as 'best'. These data need to be interpreted in the context of longer-term efficacy, effectiveness data, safety, posology, and drug acquisition costs when making treatment decisions.

DOI: https://doi.org/10.1111/bjd.19325
J Immunol. 2020 Jun 24. pii: ji1900860. [Epub ahead of print]

Interplay between RAGE and TLR4 Regulates HMGB1-Induced Inflammation by Promoting Cell Surface Expression of RAGE and TLR4.

Hanhui Zhong, Xiaolian Li, Shuangnan Zhou, Ping Jiang, Xiaolei Liu, Mingwen Ouyang, Ying Nie, Xinying Chen, Liangqing Zhang, Youtan Liu, Tao Tao, Jing Tang.

Receptor for advanced glycation end-products (RAGE) and TLR4 play an important role in the inflammatory response against High-mobility group box 1 protein (HMGB1), a late proinflammatory cytokine and a damage-associated molecular pattern. As cell surface receptors, both RAGE and TLR4 are constantly trafficking between the cytoplasm and plasma membrane. However, whether TLR4 is related to the intracellular transport of RAGE in HMGB1-induced inflammation remains unknown. In this study, we demonstrated that HMGB1 not only increased RAGE expression in both the cytoplasm and plasma membrane but also upregulated the expression of TLR4 in the plasma membrane. Knocking out of RAGE led to decreased MAPK activation, TLR4 cellular membrane expression, and corresponding inflammatory cytokine generation. Meanwhile, inhibiting MAPK activation also decreased TLR4 surface expression. These results indicated that HMGB1 may bind to cell surface RAGE receptors on the cell surface, leading to MAPK activation, thus promoting TLR4 translocation on the cell surface, but does not regulate its transcription and translation. In contrast, TLR4 can increase the transcription and translation of RAGE, which translocates to the cell surface and is able to bind to more HMGB1. The cell surface receptors TLR4 and RAGE bind to HMGB1, leading to the transcription and secretion of inflammatory cytokines. Finally, we also observed these results in the mice pseudofracture model, which is closely related to HMGB1-induced inflammatory response. All these results demonstrated that the interplay between RAGE and TLR4 are critical for HMGB1-induced inflammatory response.

DOI: https://doi.org/10.4049/jimmunol.1900860
Cell. 2020 Jun 25. pii: S0092-8674(20)30558-4. [Epub ahead of print]181(7): 1696-1696.e1

SnapShot: Jak-STAT Signaling II.

Alejandro V Villarino, Massimo Gadina, John J O'Shea, Yuka Kanno.

The JAK-STAT pathway is an evolutionarily conserved signal transduction paradigm, providing mechanisms for rapid receptor-to-nucleus communication and transcription control. Discoveries in this field provided insights into primary immunodeficiencies, inherited autoimmune and autoinflammatory diseases, and hematologic and oncologic disorders, giving rise to a new class of drugs, JAK inhibitors (or Jakinibs).

DOI: https://doi.org/10.1016/j.cell.2020.04.052
Cell Death Differ. 2020 Jun 22.

SIX1 cooperates with RUNX1 and SMAD4 in cell fate commitment of Müllerian duct epithelium.

Jumpei Terakawa, Vanida A Serna, Devi M Nair, Shigeru Sato, Kiyoshi Kawakami, Sally Radovick, Pascal Maire, Takeshi Kurita.

During female mammal reproductive tract development, epithelial cells of the lower Müllerian duct are committed to become stratified squamous epithelium of the vagina and ectocervix, when the expression of ΔNp63 transcription factor is induced by mesenchymal cells. The absence of ΔNp63 expression leads to adenosis, the putative precursor of vaginal adenocarcinoma. Our previous studies with genetically engineered mouse models have established that fibroblast growth factor (FGF)/mitogen-activated protein kinase (MAPK), bone morphogenetic protein (BMP)/SMAD, and activin A/runt-related transcription factor 1 (RUNX1) signaling pathways are independently required for ΔNp63 expression in Müllerian duct epithelium (MDE). Here, we report that sine oculis homeobox homolog 1 (SIX1) plays a critical role in the activation of ΔNp63 locus in MDE as a downstream transcription factor of mesenchymal signals. In the developing mouse reproductive tract, SIX1 expression was restricted to MDE within the future cervix and vagina. SIX1 expression was totally absent in SMAD4 null MDE and was reduced in RUNX1 null and FGFR2 null MDE, indicating that SIX1 is under the control of vaginal mesenchymal factors: BMP4, activin A and FGF7/10. Furthermore, Six1, Runx1, and Smad4 gene-dose-dependently activated ΔNp63 expression in MDE within the vaginal fornix. Using a mouse model of diethylstilbestrol (DES)-associated vaginal adenosis, we found DES action through epithelial estrogen receptor α (ESR1) inhibits activation of ΔNp63 locus in MDE by transcriptionally repressing SIX1 and RUNX1 in the vaginal fornix.

DOI: https://doi.org/10.1038/s41418-020-0579-z