OX40L blockade protects against inflammation-driven fibrosis

Treatment for fibrosis represents a critical unmet need, because fibrosis is the leading cause of death in industrialized countries, and there is no effective therapy to counteract the fibrotic process. The development of fibrosis relates to the interplay between vessel injury, immune cell activation, and fibroblast stimulation, which can occur in various tissues. Immunotherapies have provided a breakthrough in the treatment of immune diseases. The glycoprotein OX40–OX40 ligand (OX40L) axis offers the advantage of a targeted approach to costimulatory signals with limited impact on the whole immune response. Using systemic sclerosis (SSc) as a prototypic disease, we report compelling evidence that blockade of OX40L is a promising strategy for the treatment of inflammation-driven fibrosis. OX40L is overexpressed in the fibrotic skin and serum of patients with SSc, particularly in patients with diffuse cutaneous forms. Soluble OX40L was identified as a promising serum biomarker to predict the worsening of lung and skin fibrosis, highlighting the role of this pathway in fibrosis. In vivo, OX40L blockade prevents inflammation-driven skin, lung, and vessel fibrosis and induces the regression of established dermal fibrosis in different complementary mouse models. OX40L exerts potent profibrotic effects by promoting the infiltration of inflammatory cells into lesional tissues and therefore the release of proinflammatory mediators, thereafter leading to fibroblast activation.

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Prevention of NF-κB activation in vivo by a cell-permeable NF-κB inhibitor peptide

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00164.2005 ◽

2005 ◽

Vol 289 (4) ◽

pp. L536-L544 ◽

Cited By ~ 16

Author(s):

Ana L. Mora ◽

John LaVoy ◽

Martha McKean ◽

Arlene Stecenko ◽

Kenneth L. Brigham ◽

...

Keyword(s):

Recombinant Protein ◽

Stress Responses ◽

Cell Activation ◽

Immune Cell ◽

Nuclear Translocation ◽

Inflammatory Diseases ◽

New Approach ◽

Immune Cell Activation ◽

A Cell

The NF-κB/Rel transcription factor family plays a central role in coordinating the expression of a variety of genes that regulate stress responses, immune cell activation, apoptosis, proliferation, differentiation, and oncogenic transformation. Interventions that target the NF-κB pathway may be therapeutic for a variety of pathologies, especially immune/inflammatory diseases. Using membrane translocating sequence (MTS) technology, we developed a cell-permeable dominant inhibitor of NF-κB activation, termed IκBα-(ΔN)-MTS. This molecule contains a 12-amino acid MTS motif attached to the COOH-terminal region of a nondegradable inhibitor protein [IκBα-(ΔN)]. The recombinant protein enters cells and localizes in the cytoplasm. Delivery of the IκBα-(ΔN)-MTS to cell lines and primary cells inhibited nuclear translocation of NF-κB proteins induced by cell activation. The protein also effectively inhibited NF-κB activation in vivo in two different animal models: NF-κB activation in response to skin wounding in mice and NF-κB activation in lungs after endotoxin treatment in sheep. Inhibition of NF-κB by the IκBα-(ΔN)-MTS in the endotoxin model attenuated physiological responses to endotoxemia. These data demonstrate that activation of NF-κB can be inhibited using a recombinant protein designed to penetrate into cells. This technology may provide a new approach to NF-κB pathway-targeted therapies.

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Innate immune cell activation and epigenetic remodeling in symptomatic and asymptomatic atherosclerosis in humans in vivo

Atherosclerosis ◽

10.1016/j.atherosclerosis.2016.10.019 ◽

2016 ◽

Vol 254 ◽

pp. 228-236 ◽

Cited By ~ 70

Author(s):

Siroon Bekkering ◽

Inge van den Munckhof ◽

Tim Nielen ◽

Evert Lamfers ◽

Charles Dinarello ◽

...

Keyword(s):

Cell Activation ◽

Immune Cell ◽

Innate Immune ◽

Innate Immune Cell ◽

Immune Cell Activation ◽

Epigenetic Remodeling

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SAP130 released by damaged tubule drives necroinflammation via miRNA-219c/Mincle signaling in acute kidney injury

Cell Death and Disease ◽

10.1038/s41419-021-04131-7 ◽

2021 ◽

Vol 12 (10) ◽

Author(s):

Lin-Li Lv ◽

Cui Wang ◽

Zuo-Lin Li ◽

Jing-Yuan Cao ◽

Xin Zhong ◽

...

Keyword(s):

Acute Kidney Injury ◽

Macrophage Activation ◽

Cell Activation ◽

Immune Cell ◽

Kidney Injury ◽

Novel Therapy ◽

Tubular Cells ◽

Immune Cell Activation ◽

Dependent Mechanism

AbstractTubules injury and immune cell activation are the common pathogenic mechanisms in acute kidney injury (AKI). However, the exact modes of immune cell activation following tubule damage are not fully understood. Here we uncovered that the release of cytoplasmic spliceosome associated protein 130 (SAP130) from the damaged tubular cells mediated necroinflammation by triggering macrophage activation via miRNA-219c(miR-219c)/Mincle-dependent mechanism in unilateral ureteral obstruction (UUO) and cisplatin-induced AKI mouse models, and in patients with acute tubule necrosis (ATN). In the AKI kidneys, we found that Mincle expression was tightly correlated to the necrotic tubular epithelial cells (TECs) with higher expression of SAP130, a damaged associated molecule pattern (DAMP), suggesting that SAP130 released from damaged tubular cells may trigger macrophage activation and necroinflammation. This was confirmed in vivo in which administration of SAP130-rich supernatant from dead TECs or recombinant SAP130 promoted Mincle expression and macrophage accumulation which became worsen with profound tubulointerstitial inflammation in LPS-primed Mincle WT mice but not in Mincle deficient mice. Further studies identified that Mincle was negatively regulated via miR-219c-3p in macrophages as miR-219c-3p bound Mincle 3′-UTR to inhibit Mincle translation. Besides, lentivirus-mediated renal miR-219c-3p overexpression blunted Mincle and proinflammatory cytokine expression as well as macrophage infiltration in the inflamed kidney of UUO mice. In conclusion, SAP130 is released by damaged tubules which elicit Mincle activation on macrophages and renal necroinflammation via the miR-219c-3p-dependent mechanism. Results from this study suggest that targeting miR-219c-3p/Mincle signaling may represent a novel therapy for AKI.

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OS12.4 In vivo dynamics and targeting of vessel co-option in glioma

Neuro-Oncology ◽

10.1093/neuonc/noz126.076 ◽

2019 ◽

Vol 21 (Supplement_3) ◽

pp. iii23-iii23

Author(s):

G Seano ◽

A Griveau ◽

S Shelton ◽

S Krishnan ◽

N Wang ◽

...

Keyword(s):

Single Cell ◽

Cell Activation ◽

Immune Cell ◽

Glioma Cells ◽

Angiogenic Therapy ◽

Immune Cell Activation ◽

Anti Vegf ◽

Oligodendrocyte Precursor ◽

Wnt Inhibitors

Abstract BACKGROUND Gliomas comprise heterogeneous malignant glial and stromal cells. While blood vessel co-option is a potential mechanism to escape anti-angiogenic therapy, the relevance of glial phenotype in this process is unclear. MATERIAL AND METHODS Here, we intravitally study preclinical syngenetic models of glioma as well as patient-derived cells transplanted orthotopically. Moreover, we profoundly confirm our preclinical results with histological studies on patient specimens. RESULTS We show that Olig2+ oligodendrocyte precursor-like glioma cells invade by single-cell vessel co-option and preserve the blood-brain barrier (BBB). Conversely, Olig2-negative glioma cells form dense perivascular collections and promote angiogenesis and BBB breakdown, leading to innate immune cell activation. Experimentally, Olig2 promotes Wnt7b expression, a finding that correlates in human glioma profiling. Targeted Wnt7a/7b deletion or pharmacologic Wnt inhibition blocks Olig2+ glioma single-cell vessel co-option and enhances responses to temozolomide. Finally, Olig2 and Wnt7 become upregulated after anti-VEGF treatment in preclinical models and patients. CONCLUSION Here, we show that glioma is able to employ vessel co-option, i.e. the movement of tumor cells towards and along the pre-existing vasculature. Glioma oligodendrocyte-like (OPCL) cells express Wnt7 that is necessary for vessel co-option and Wnt inhibitors significantly improve survival with temozolomide. Moreover, we demonstrated that anti-VEGF-treatment of glioma selects for Olig2/Wnt7+ cells

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Structural and Clinical Correlates of a Periventricular Gradient of Neuroinflammation in Multiple Sclerosis

Neurology ◽

10.1212/wnl.0000000000011700 ◽

2021 ◽

pp. 10.1212/WNL.0000000000011700

Author(s):

Emilie Poirion ◽

Matteo Tonietto ◽

François-Xavier Lejeune ◽

Vito A.G. Ricigliano ◽

Marine Boudot de la Motte ◽

...

Keyword(s):

Multiple Sclerosis ◽

Cell Activation ◽

Immune Cell ◽

Innate Immune ◽

Innate Immune Cell ◽

Magnetization Transfer Ratio ◽

Immune Cell Activation ◽

Microstructural Damage ◽

Ventricular Surface

Objectives:To explore in-vivo innate immune cell activation as a function of the distance from ventricular CSF in patients with Multiple Sclerosis (MS) using [18F]-DPA714 PET, and to investigate its relationship with periventricular microstructural damage, evaluated by magnetization transfer ratio (MTR), and with trajectories of disability worsening.Methods:Thirty-seven MS patients and nineteen healthy controls underwent MRI and [18F]-DPA714 TSPO dynamic PET, from which individual maps of voxels characterized by innate immune cell activation (DPA+) were generated. White matter (WM) was divided in 3mm-thick concentric rings radiating from the ventricular surface toward the cortex, and the percentage of DPA+ voxels and mean MTR were extracted from each ring. Two-year trajectories of disability worsening were collected to identify patients with and without recent disability worsening.Results:The percentage of DPA+ voxels was higher in patients compared to controls in the periventricular WM (p=6.10e-6), and declined with increasing distance from ventricular surface, with a steeper gradient in patients compared to controls (p=0.001). This gradient was found both in periventricular lesions and normal-appearing WM. In the total WM, it correlated with a gradient of microstructural tissue damage measured by MTR (rs=-0.65, p=1.0e-3). When compared to clinically stable patients, patients with disability worsening were characterized by a higher percentage of DPA+ voxels in the periventricular normal-appearing WM (p=0.025).Conclusions:Our results demonstrate that in MS the innate immune cell activation predominates in periventricular regions and associates with microstructural damage and disability worsening. This could result from the diffusion of pro-inflammatory CSF-derived factors into surrounding tissues.

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Deoxyribonuclease 1-Mediated Clearance of Circulating Chromatin Prevents From Immune Cell Activation and Pro-inflammatory Cytokine Production, a Phenomenon Amplified by Low Trap1 Activity: Consequences for Systemic Lupus Erythematosus

Frontiers in Immunology ◽

10.3389/fimmu.2021.613597 ◽

2021 ◽

Vol 12 ◽

Author(s):

Jasmin Felux ◽

Annika Erbacher ◽

Magali Breckler ◽

Roxane Hervé ◽

Delphine Lemeiter ◽

...

Keyword(s):

Immune Cells ◽

Lupus Erythematosus ◽

Cell Activation ◽

Immune Cell ◽

Wild Type ◽

Systemic Lupus ◽

Immune Cell Activation ◽

Deficient Mice

Increased concentrations of circulating chromatin, especially oligo-nucleosomes, are observed in sepsis, cancer and some inflammatory autoimmune diseases like systemic lupus erythematosus (SLE). In SLE, circulating nucleosomes mainly result from increased apoptosis and decreased clearance of apoptotic cells. Once released, nucleosomes behave both as an autoantigen and as a damage-associated molecular pattern (DAMP) by activating several immune cells, especially pro-inflammatory cells. Deoxyribonuclease 1 (DNase1) is a major serum nuclease whose activity is decreased in mouse and human lupus. Likewise, the mitochondrial chaperone tumor necrosis factor (TNF) receptor-associated protein-1 (Trap1) protects against oxidative stress, which is increased in SLE. Here, using wild type, DNase1-deficient and DNase1/Trap1-deficient mice, we demonstrate that DNase1 is a major serum nuclease involved in chromatin degradation, especially when the plasminogen system is activated. In vitro degradation assays show that chromatin digestion is strongly impaired in serum from DNase1/Trap1-deficient mice as compared to wild type mice. In vivo, after injection of purified chromatin, clearance of circulating chromatin is delayed in DNase1/Trap1-deficient mice in comparison to wild type mice. Since defective chromatin clearance may lead to chromatin deposition in tissues and subsequent immune cell activation, spleen cells were stimulated in vitro with chromatin. Splenocytes were activated by chromatin, as shown by interleukin (IL)-12 secretion and CD69 up-regulation. Moreover, cell activation was exacerbated when Trap1 is deficient. Importantly, we also show that cytokines involved in lupus pathogenesis down-regulate Trap1 expression in splenocytes. Therefore, combined low activities of both DNase1 and Trap1 lead to an impaired degradation of chromatin in vitro, delayed chromatin clearance in vivo and enhanced activation of immune cells. This situation may be encountered especially, but not exclusively, in SLE by the negative action of cytokines on Trap1 expression.

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685 A highly selective and potent HPK1 inhibitor enhances immune cell activation and induces robust tumor growth inhibition in a murine syngeneic tumor model

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0685 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A724-A724

Author(s):

David Ciccone ◽

Vad Lazari ◽

Ian Linney ◽

Michael Briggs ◽

Samantha Carreiro ◽

...

Keyword(s):

T Cells ◽

Cell Activation ◽

Immune Cell ◽

Tumor Model ◽

Tumor Growth Inhibition ◽

Immune Cell Activation ◽

Syngeneic Tumor ◽

Human T Cells

BackgroundHPK1, a member of the MAP4K family of protein serine/threonine kinases, is involved in regulating signal transduction cascades in cells of hematopoietic origin. Recent data from HPK1 knockout animals and kinase-inactive knock-in animals underscores the role of HPK1 in negatively regulating immune cell activation. This negative-feedback role of HPK1 combined with its restricted expression in cells of hematopoietic origin, make it a compelling drug target for enhancing anti-tumor immunity.MethodsA structure-based drug design approach was used to identify potent and selective inhibitors of HPK1. Biochemical assays, as well as primary human and mouse immune cell-based activation assays, were utilized for multiple iterations of structure-activity relationship (SAR) studies. In vivo efficacy, target engagement and pharmacodynamic data were generated using murine syngeneic tumor models.ResultsA highly potent, HPK1 inhibitor was identified, that showed high selectivity against T cell-specific kinases and kinases in the MAP4K family. In vitro, HPK1 small molecule inhibition resulted in enhanced IL-2 production in primary mouse and human T cells, enhanced IL-6 and IgG production in primary human B cells, and enhanced mouse dendritic cell activation and antigen presentation capacity. Furthermore, HPK1 inhibition alleviated the immuno-suppressive effects of PGE2 on naïve human T cells and restored the proliferative capacity of exhausted human T cells. In vivo, HPK1 inhibitionHPK1 inhibition abrogated T cell receptor-stimulated phospho-SLP-76, enhanced cytokine production, and mediated robust tumor growth inhibition in a murine syngeneic tumor model.ConclusionsPharmacological blockade of HPK1 kinase activity represents a novel and potentially valuable immunomodulatory approach for anti-tumor immunity.

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Innate immune cell activation in symptomatic and asymptomatic atherosclerosis in humans in vivo

Atherosclerosis ◽

10.1016/j.atherosclerosis.2016.07.077 ◽

2016 ◽

Vol 252 ◽

pp. e256

Author(s):

S. Bekkering ◽

I. van den Munckhof ◽

T. Nielen ◽

J. Rutten ◽

J. de Graaf ◽

...

Keyword(s):

Cell Activation ◽

Immune Cell ◽

Innate Immune ◽

Innate Immune Cell ◽

Immune Cell Activation

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The cytosolic DNA sensor cGAS recognizes neutrophil extracellular traps

Science Signaling ◽

10.1126/scisignal.aax7942 ◽

2021 ◽

Vol 14 (673) ◽

pp. eaax7942

Author(s):

Falko Apel ◽

Liudmila Andreeva ◽

Lorenz Sebastian Knackstedt ◽

Robert Streeck ◽

Christian Karl Frese ◽

...

Keyword(s):

Cell Activation ◽

Immune Cell ◽

Myeloid Cells ◽

Neutrophil Extracellular Traps ◽

Type I Interferons ◽

Type I ◽

Immune Cell Activation ◽

Extracellular Traps ◽

Regulated Cell Death

Neutrophil extracellular traps (NETs) are structures consisting of chromatin and antimicrobial molecules that are released by neutrophils during a form of regulated cell death called NETosis. NETs trap invading pathogens, promote coagulation, and activate myeloid cells to produce type I interferons (IFNs), proinflammatory cytokines that regulate the immune system. Here, we showed that macrophages and other myeloid cells phagocytosed NETs. Once in phagosomes, NETs translocated to the cytosol, where the DNA backbones of these structures activated the innate immune sensor cyclic GMP-AMP synthase (cGAS) and induced type I IFN production. The NET-associated serine protease neutrophil elastase (NE) mediated the activation of this pathway. We showed that NET induction in mice treated with the lectin concanavalin A, a model of autoimmune hepatitis, resulted in cGAS-dependent stimulation of an IFN response, suggesting that NETs activated cGAS in vivo. Thus, our findings suggest that cGAS is a sensor of NETs, mediating immune cell activation during infection.

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