scholarly journals POS0428 REGULATION OF IFN SIGNATURE BY HDAC CLASS IIa-CD52 AXIS IN SYSTEMIC SCLEROSIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 442.2-443
Author(s):  
M. Rudnik ◽  
A. Hukara ◽  
P. Blyszczuk ◽  
O. Distler ◽  
G. Kania
Keyword(s):  
Valproic Acid ◽  
Systemic Sclerosis ◽  
Pathway Analysis ◽  
Histone Deacetylases ◽  
Molecular Mechanisms ◽  
Cell Receptor ◽  
Type I ◽  
Illumina Hiseq ◽  
Induced Expression ◽  
Protein Levels

Background:Systemic sclerosis (SSc) is associated with an interferon (IFN) signature, which is defined by a higher expression of IFN-stimulated genes (mainly in response to IFNα). Histone deacetylases (HDACs) are a family of epigenetic modifiers mediating immune function. HDACs function via diverse molecular mechanisms, including direct inhibition of gene transcription or indirectly through modulation of nuclear transcription factors such as NF-κB and STATs. CD52 protein regulates T cell receptor and NF-κB signalling. Previously, we showed downregulation of CD52 in SSc monocytes, however, the influence of CD52 on IFN signalling has not been studied yet.Objectives:We investigated the role of CD52 in the regulation of IFN response in monocytes. Moreover, we explored the regulatory mechanisms of CD52 expression to identify the involvement of HDACs in that process.Methods:RNAseq of CD14+ monocytes isolated from peripheral blood of lcSSc (n=5, age=54.4±6.7), dcSSc patients (n=5, age=51.8±7.2) and age- and sex-matched healthy controls (HC) (n=5, age=50.8±9.7) was performed using Illumina HiSeq 4000 platform. Differentially expressed genes were computed using DeSEQ2 algorithm. Gene ontology and pathway analysis were performed using Metacore software and ShinyApp. CD52 activity in monocytes was blocked by monoclonal antibody Alemtuzumab [10ug/ml] and IFN signature genes expression was assessed upon IFNα stimulation [1ng/ml] by qPCR and ELISA. HDAC-dependent regulation of CD52 expression in CD14+ monocytes from HC was analysed on mRNA and protein levels after treatment with pan-HDAC inhibitor valproic acid and HDAC class IIa inhibitor TMP269 [both 2.5μM].Results:Pathway analysis revealed significant alterations in interferon signalling in SSc monocytes. Monocytes treated with Alemtuzumab and IFNα showed induced expression of STAT1 (p=0.023, N=4), CXCL9 (p=0.062, N=4) and CXCL10 (p=0.005, N=4). RNAseq revealed a specific HDAC expression pattern in SSc monocytes with induced expression of HDAC4, 6, 10, 11 (p<0.05) and reduced HDAC1, 3 and 8 (p<0.05). CD52 mRNA was significantly decreased after IFNα stimulation (p=0.001, N=3). Treatment with valproic acid and HDAC class IIa inhibitor TMP269 resulted in decreased phosphorylation of STAT1 (p<0.01, N=4) followed by a declined level of CXCL10 (p<0.01, N=4) and restored level of CD52 mRNA (p<0.001, N=4).Conclusion:Our findings demonstrated a new aspect of pro-inflammatory type I IFN signalling in SSc. We described a novel regulation feedback loop in monocytes, in which CD52 suppresses IFN signature, while its expression is inhibited by IFN-induced HDAC activity (mainly HDAC class IIa). Therefore, targeting the CD52-IFN-HDAC axis might serve as a novel therapeutic strategy in SSc.Disclosure of Interests:Michal Rudnik: None declared, Amela Hukara: None declared, Przemyslaw Blyszczuk: None declared, Oliver Distler Speakers bureau: Actelion, Bayer, Boehringer Ingelheim, Medscape, Novartis, Roche, Menarini, Mepha, MSD, iQone, Novartis, Pfizer, Consultant of: Abbvie, Actelion, Acceleron Pharma, Amgen, AnaMar, Arxx Therapeutics, Bayer, Baecon Discovery, Blade Therapeutics, Boehringer, CSL Behring, ChemomAb, Corpuspharma, Curzion Pharmaceuticals, Ergonex, Galapagos NV, GSK, Glenmark Pharmaceuticals, Inventiva, Italfarmaco, iQvia, Kymera, Medac, Medscape, Mitsubishi Tanabe Pharma, MSD, Roche, Sanofi, UCB, Lilly, Target BioScience, Pfizer, Grant/research support from: Actelion, Bayer, Boehringer Ingelheim, Kymera Therapeutics, Mitsubishi Tanabe, Gabriela Kania: None declared

scholarly journals Co-ordinate increase in the expression of type I and type III collagen genes in progressive systemic sclerosis fibroblasts

1986 ◽  
Vol 237 (3) ◽  
pp. 837-843 ◽  
Author(s):  
S A Jimenez ◽  
G Feldman ◽  
R I Bashey ◽  
R Bienkowski ◽  
J Rosenbloom
Keyword(s):  
Systemic Sclerosis ◽  
Cell Lines ◽  
Molecular Mechanisms ◽  
Dermal Fibroblast ◽  
Blot Hybridization ◽  
Progressive Systemic Sclerosis ◽  
Dermal Fibroblasts ◽  
Type Iii Collagen ◽  
Type I ◽  
Dot Blot

Progressive systemic sclerosis (PSS), is a connective tissue disease characterized by excessive accumulation of collagen in the skin and various internal organs which is due, at least in part, to increased collagen production by PSS fibroblasts. In order to examine the molecular mechanisms responsible for this abnormality, we compared the kinetics of collagen biosynthesis, the intracellular degradation of collagen and the expression of Types I and III procollagen genes between normal and PSS dermal fibroblasts in culture. Two age- and sex-matched normal and PSS dermal fibroblast cell lines were studied. The results showed that the PSS cultures produced higher amounts of collagen than did normal fibroblasts and displayed an abnormal kinetic pattern. Furthermore, the PSS cells showed a slight but statistically significant increase in the fraction of collagen degraded intracellularly when compared with normal cells (23% against 18% respectively). The levels of mRNA for procollagen Types I and III were determined by Northern and dot-blot hybridization with specific cloned cDNA probes for alpha 1(I), alpha 2(I) and alpha 1(III) and it was found that they were 2-3-fold higher for each of the three chains in the PSS cell lines compared with the controls. These findings indicate, therefore, that the overproduction of collagen characteristic of PSS fibroblasts can be largely accounted for by the increased levels of collagen mRNA.

scholarly journals Human Metapneumovirus Induces IRF1 via TANK-Binding Kinase 1 and Type I IFN

2021 ◽  
Vol 12 ◽  
Author(s):  
Simon Loevenich ◽  
Alix S. Spahn ◽  
Kristin Rian ◽  
Victor Boyartchuk ◽  
Marit Walbye Anthonsen
Keyword(s):  
Fungal Pathogens ◽  
Molecular Mechanisms ◽  
Viral Infections ◽  
Human Monocyte ◽  
Cell Types ◽  
Human Metapneumovirus ◽  
Type I ◽  
Protein Levels ◽  
Antiviral Responses ◽  
Mucosal Surfaces

The innate immune and host-protective responses to viruses, such as the airway pathogen human metapneumovirus (HMPV), depend on interferons (IFNs) that is induced through TANK-binding kinase 1 (TBK1) and IFN regulatory factors (IRFs). The transcription factor IRF1 is important for host resistance against several viruses and has a key role in induction of IFN-λ at mucosal surfaces. In most cell types IRF1 is expressed at very low levels, but its mRNA is rapidly induced when the demand for IRF1 activity arises. Despite general recognition of the importance of IRF1 to antiviral responses, the molecular mechanisms by which IRF1 is regulated during viral infections are not well understood. Here we identify the serine/threonine kinase TBK1 and IFN-β as critical regulators of IRF1 mRNA and protein levels in human monocyte-derived macrophages. We find that inhibition of TBK1 activity either by the semi-selective TBK1/IKKε inhibitor BX795 or by siRNA-mediated knockdown abrogates HMPV-induced expression of IRF1. Moreover, we show that canonical NF-κB signaling is involved in IRF1 induction and that the TBK1/IKKε inhibitor BX795, but not siTBK1 treatment, impairs HMPV-induced phosphorylation of the NF-κB subunit p65. At later time-points of the infection, IRF1 expression depended heavily on IFN-β-mediated signaling via the IFNAR-STAT1 pathway. Hence, our results suggest that TBK1 activation and TBK1/IKKε-mediated phosphorylation of the NF-κB subunit p65 control transcription of IRF1. Our study identifies a novel mechanism for IRF1 induction in response to viral infection of human macrophages that could be relevant not only to defense against HMPV, but also to other viral, bacterial and fungal pathogens.

scholarly journals Valproic Acid and Epilepsy: From Molecular Mechanisms to Clinical Evidences

2019 ◽  
Vol 17 (10) ◽  
pp. 926-946 ◽  
Author(s):  
Michele Romoli ◽  
Petra Mazzocchetti ◽  
Renato D'Alonzo ◽  
Sabrina Siliquini ◽  
Victoria Elisa Rinaldi ◽  
...  
Keyword(s):  
Valproic Acid ◽  
Broad Spectrum ◽  
Histone Deacetylases ◽  
Molecular Mechanisms ◽  
Neurological Diseases ◽  
Ionic Currents ◽  
Convulsive Status Epilepticus ◽  
Drug Of Choice ◽  
Positive Modulator ◽  
Synaptic Effects

After more than a century from its discovery, valproic acid (VPA) still represents one of the most efficient antiepileptic drugs (AEDs). Pre and post-synaptic effects of VPA depend on a very broad spectrum of actions, including the regulation of ionic currents and the facilitation of GABAergic over glutamatergic transmission. As a result, VPA indirectly modulates neurotransmitter release and strengthens the threshold for seizure activity. However, even though participating to the anticonvulsant action, such mechanisms seem to have minor impact on epileptogenesis. Nonetheless, VPA has been reported to exert anti-epileptogenic effects. Epigenetic mechanisms, including histone deacetylases (HDACs), BDNF and GDNF modulation are pivotal to orientate neurons toward a neuroprotective status and promote dendritic spines organization. From such broad spectrum of actions comes constantly enlarging indications for VPA. It represents a drug of choice in child and adult with epilepsy, with either general or focal seizures, and is a consistent and safe IV option in generalized convulsive status epilepticus. Moreover, since VPA modulates DNA transcription through HDACs, recent evidences point to its use as an anti-nociceptive in migraine prophylaxis, and, even more interestingly, as a positive modulator of chemotherapy in cancer treatment. Furthermore, VPA-induced neuroprotection is under investigation for benefit in stroke and traumatic brain injury. Hence, VPA has still got its place in epilepsy, and yet deserves attention for its use far beyond neurological diseases. In this review, we aim to highlight, with a translational intent, the molecular basis and the clinical indications of VPA.

scholarly journals Identification of biomarkers associated with metabolic cardiovascular disease using mRNA-SNP-miRNA regulatory network analysis

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Zhiyuan Fan ◽  
Wenjuan Peng ◽  
Zhiwen Wang ◽  
Ling Zhang ◽  
Kuo Liu
Keyword(s):  
Cardiovascular Disease ◽  
Molecular Mechanisms ◽  
Early Stage ◽  
Expression Profiles ◽  
Interaction Network ◽  
Cell Receptor ◽  
Differentially Expressed ◽  
Eqtl Analysis ◽  
Nucleotide Polymorphisms ◽  
Illumina Hiseq

Abstract Background CVD is the leading cause of death in T2DM patients. However, few biomarkers have been identified to detect and diagnose CVD in the early stage of T2DM. The aim of our study was to identify the important mRNAs, micro (mi)RNAs and SNPs (single nucleotide polymorphisms) that are associated with metabolic cardiovascular disease. Materials and methods Expression profiles and GWAS data were obtained from Gene Expression Omnibus (GEO) database. MiRNA-sequencing was conducted by Illumina HiSeq 2000 platform in T2DM patients and T2DM with CVD patients. EQTL analysis and gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted. MRNA-miRNA co-expression network and mRNA-SNP-miRNA interaction network were established and visualized by Cytoscape 3.7.2. Results In our study, we identified 56 genes and 16 miRNAs that were significantly differentially expressed. KEGG analyses results indicated that B cell receptor signaling pathway and hematopoietic cell lineage were included in the biological functions of differentially expressed genes. MRNA-miRNA co-expression network and mRNA-SNP-miRNA interaction network illustrated that let-7i-5p, RASGRP3, KRT1 and CEP41 may be potential biomarkers for the early detection and diagnosis of CVD in T2DM patients. Conclusion Our results suggested that downregulated let-7i-5p, and upregulated RASGRP3, KRT1 and CEP41 may play crucial roles in molecular mechanisms underlying the initiation and development of CVD in T2DM patients.

Identification of biomarkers associated with metabolic cardiovascular disease using mRNA-SNP-miRNA regulatory network analysis

2021 ◽  
Author(s):  
Zhiyuan Fan ◽  
Wenjuan Peng ◽  
Zhiwen Wang ◽  
ling Zhang ◽  
Kuo Liu
Keyword(s):  
Cardiovascular Disease ◽  
Molecular Mechanisms ◽  
Early Stage ◽  
Expression Profiles ◽  
Interaction Network ◽  
Cell Receptor ◽  
Differentially Expressed ◽  
Eqtl Analysis ◽  
Nucleotide Polymorphisms ◽  
Illumina Hiseq

Abstract Background: CVD is the leading cause of death in T2DM patients. However, few biomarkers have been identified to detect and diagnose CVD in the early stage of T2DM. The aim of our study was to identify the important mRNAs, micro (mi)RNAs and SNPs (single nucleotide polymorphisms) that are associated with metabolic cardiovascular disease. Materials and methods: Expression profiles and GWAS data were obtained from Gene Expression Omnibus (GEO) database. MiRNA-sequencing was conducted by Illumina HiSeq 2000 platform in T2DM patients and T2DM with CVD patients. EQTL analysis and gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted. MRNA-miRNA co-expression network and mRNA-SNP-miRNA interaction network were established and visualized by Cytoscape 3.7.2.Results: In our study, we identified 56 genes and 16 miRNAs that were significantly differentially expressed. GO and KEGG analyses results indicated that B cell receptor signaling pathway and hematopoietic cell lineage were included in the biological functions of differentially expressed genes. MRNA-miRNA co-expression network and mRNA-SNP-miRNA interaction network illustrated that let-7i-5p, RASGRP3, KRT1 and CEP41 may be potential biomarkers for the early detection and diagnosis of CVD in T2DM patients.Conclusion: Our results suggested that downregulated let-7i-5p, and upregulated RASGRP3, KRT1 and CEP41 may play crucial roles in molecular mechanisms underlying the initiation and development of CVD in T2DM patients.

Classic and Novel Signaling Pathways Involved in Cancer: Targeting the NF-κB and Syk Signaling Pathways

2019 ◽  
Vol 14 (3) ◽  
pp. 219-225 ◽  
Author(s):  
Cong Tang ◽  
Guodong Zhu
Keyword(s):  
Cancer Therapy ◽  
Tyrosine Kinase ◽  
Signaling Pathways ◽  
Molecular Mechanisms ◽  
Therapeutic Potential ◽  
Cell Receptor ◽  
Transmembrane Receptors ◽  
Biological Responses ◽  
Different Types

The nuclear factor kappa B (NF-κB) consists of a family of transcription factors involved in the regulation of a wide variety of biological responses. Growing evidence support that NF-κB plays a major role in oncogenesis as well as its well-known function in the regulation of immune responses and inflammation. Therefore, we made a review of the diverse molecular mechanisms by which the NF-κB pathway is constitutively activated in different types of human cancers and the potential role of various oncogenic genes regulated by this transcription factor in cancer development and progression. We also discussed various pharmacological approaches employed to target the deregulated NF-κB signaling pathway and their possible therapeutic potential in cancer therapy. Moreover, Syk (Spleen tyrosine kinase), non-receptor tyrosine kinase which mediates signal transduction downstream of a variety of transmembrane receptors including classical immune-receptors like the B-cell receptor (BCR), which can also activate the inflammasome and NF-κB-mediated transcription of chemokines and cytokines in the presence of pathogens would be discussed as well. The highlight of this review article is to summarize the classic and novel signaling pathways involved in NF-κB and Syk signaling and then raise some possibilities for cancer therapy.

scholarly journals The AP-1 Transcription Factor Fosl-2 Regulates Autophagy in Cardiac Fibroblasts during Myocardial Fibrogenesis

2021 ◽  
Vol 22 (4) ◽  
pp. 1861
Author(s):  
Jemima Seidenberg ◽  
Mara Stellato ◽  
Amela Hukara ◽  
Burkhard Ludewig ◽  
Karin Klingel ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Molecular Mechanisms ◽  
Cardiac Fibrosis ◽  
Type I Collagen ◽  
Transforming Growth Factor ◽  
Cardiac Fibroblasts ◽  
Smooth Muscle Actin ◽  
Beclin 1 ◽  
Type I ◽  
Alpha Smooth Muscle Actin

Background: Pathological activation of cardiac fibroblasts is a key step in development and progression of cardiac fibrosis and heart failure. This process has been associated with enhanced autophagocytosis, but molecular mechanisms remain largely unknown. Methods and Results: Immunohistochemical analysis of endomyocardial biopsies showed increased activation of autophagy in fibrotic hearts of patients with inflammatory cardiomyopathy. In vitro experiments using mouse and human cardiac fibroblasts confirmed that blockade of autophagy with Bafilomycin A1 inhibited fibroblast-to-myofibroblast transition induced by transforming growth factor (TGF)-β. Next, we observed that cardiac fibroblasts obtained from mice overexpressing transcription factor Fos-related antigen 2 (Fosl-2tg) expressed elevated protein levels of autophagy markers: the lipid modified form of microtubule-associated protein 1A/1B-light chain 3B (LC3BII), Beclin-1 and autophagy related 5 (Atg5). In complementary experiments, silencing of Fosl-2 with antisense GapmeR oligonucleotides suppressed production of type I collagen, myofibroblast marker alpha smooth muscle actin and autophagy marker Beclin-1 in cardiac fibroblasts. On the other hand, silencing of either LC3B or Beclin-1 reduced Fosl-2 levels in TGF-β-activated, but not in unstimulated cells. Using a cardiac hypertrophy model induced by continuous infusion of angiotensin II with osmotic minipumps, we confirmed that mice lacking either Fosl-2 (Ccl19CreFosl2flox/flox) or Atg5 (Ccl19CreAtg5flox/flox) in stromal cells were protected from cardiac fibrosis. Conclusion: Our findings demonstrate that Fosl-2 regulates autophagocytosis and the TGF-β-Fosl-2-autophagy axis controls differentiation of cardiac fibroblasts. These data provide a new insight for the development of pharmaceutical targets in cardiac fibrosis.

scholarly journals Listeria exploits IFITM3 to suppress antibacterial activity in phagocytes

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Joel M. J. Tan ◽  
Monica E. Garner ◽  
James M. Regeimbal ◽  
Catherine J. Greene ◽  
Jorge D. Rojas Márquez ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Molecular Mechanisms ◽  
Immune Cell ◽  
Transmembrane Protein ◽  
Foodborne Pathogen ◽  
Systemic Infection ◽  
Cytokine Signaling ◽  
Type I ◽  
Type I Ifn ◽  
Antiviral Protein

AbstractThe type I interferon (IFN) signaling pathway has important functions in resistance to viral infection, with the downstream induction of interferon stimulated genes (ISG) protecting the host from virus entry, replication and spread. Listeria monocytogenes (Lm), a facultative intracellular foodborne pathogen, can exploit the type I IFN response as part of their pathogenic strategy, but the molecular mechanisms involved remain unclear. Here we show that type I IFN suppresses the antibacterial activity of phagocytes to promote systemic Lm infection. Mechanistically, type I IFN suppresses phagosome maturation and proteolysis of Lm virulence factors ActA and LLO, thereby promoting phagosome escape and cell-to-cell spread; the antiviral protein, IFN-induced transmembrane protein 3 (IFITM3), is required for this type I IFN-mediated alteration. Ifitm3−/− mice are resistant to systemic infection by Lm, displaying decreased bacterial spread in tissues, and increased immune cell recruitment and pro-inflammatory cytokine signaling. Together, our findings show how an antiviral mechanism in phagocytes can be exploited by bacterial pathogens, and implicate IFITM3 as a potential antimicrobial therapeutic target.

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