scholarly journals IL-1ra Secreted by ATP-Induced P2Y2Negatively Regulates MUC5AC Overproduction via PLCβ3 during Airway Inflammation

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Jee-Yeong Jeong ◽  
Jiwook Kim ◽  
Bokyoum Kim ◽  
Joowon Kim ◽  
Yusom Shin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Expression ◽  
Airway Inflammation ◽  
Respiratory Diseases ◽  
Inflammatory Diseases ◽  
Dominant Negative ◽  
Regulatory Effect ◽  
Regulatory Pathway ◽  
Cytokines And Chemokines ◽  
Dominant Negative Mutation

Mucus secretion is often uncontrolled in many airway inflammatory diseases of humans. Identifying the regulatory pathway(s) of mucus gene expression, mucus overproduction, and hypersecretion is important to alleviate airway inflammation in these diseases. However, the regulatory signaling pathway controlling mucus overproduction has not been fully identified yet. In this study, we report that the ATP/P2Y2complex secretes many cytokines and chemokines to regulate airway inflammation, among which IL-1 receptor antagonist (IL-1ra) downregulatesMUC5ACgene expression via the inhibition of Gαq-induced Ca2+signaling. IL-1ra inhibited IL-1αprotein expression and secretion, and vice versa. Interestingly, ATP/P2Y2-induced IL-1ra and IL-1αsecretion were both mediated by PLCβ3. A dominant-negative mutation in the PDZ-binding domain of PLCβ3 inhibited ATP/P2Y2-induced IL-1ra and IL-1αsecretion. IL-1αin the presence of the ATP/P2Y2complex activated the ERK1/2 pathway in a greater degree and for a longer duration than the ATP/P2Y2complex itself, which was dramatically inhibited by IL-1ra. These findings suggest that secreted IL-1ra exhibits a regulatory effect on ATP/P2Y2-inducedMUC5ACgene expression, through inhibition of IL-1αsecretion, to maintain the mucus homeostasis in the airway. Therefore, IL-1ra could be an excellent modality for regulating inflamed airway microenvironments in respiratory diseases.

scholarly journals An Autocrine Wnt5a Loop Promotes NF-κB Pathway Activation and Cytokine/Chemokine Secretion in Melanoma

Cells ◽  
2019 ◽  
Vol 8 (9) ◽  
pp. 1060 ◽  
Author(s):  
Gastón Barbero ◽  
María Victoria Castro ◽  
María Belén Villanueva ◽  
María Josefina Quezada ◽  
Natalia Brenda Fernández ◽  
...  
Keyword(s):  
Nuclear Translocation ◽  
Epithelial Mesenchymal Transition ◽  
Inflammatory Responses ◽  
Inflammatory Diseases ◽  
Melanoma Cells ◽  
Dominant Negative ◽  
Migration And Invasion ◽  
Soluble Receptor ◽  
Mesenchymal Transition ◽  
Cytokines And Chemokines

Wnt5a signaling has been implicated in the progression of cancer by regulating multiple cellular processes, largely migration and invasion, epithelial-mesenchymal transition (EMT), and metastasis. Since Wnt5a signaling has also been involved in inflammatory processes in infectious and inflammatory diseases, we addressed the role of Wnt5a in regulating NF-κB, a pivotal mediator of inflammatory responses, in the context of cancer. The treatment of melanoma cells with Wnt5a induced phosphorylation of the NF-κB subunit p65 as well as IKK phosphorylation and IκB degradation. By using cDNA overexpression, RNA interference, and dominant negative mutants we determined that ROR1, Dvl2, and Akt (from the Wnt5a pathway) and TRAF2 and RIP (from the NF-κB pathway) are required for the Wnt5a/NF-κB crosstalk. Wnt5a also induced p65 nuclear translocation and increased NF-κB activity as evidenced by reporter assays and a NF-κB-specific upregulation of RelB, Bcl-2, and Cyclin D1. Further, stimulation of melanoma cells with Wnt5a increased the secretion of cytokines and chemokines, including IL-6, IL-8, IL-11, and IL-6 soluble receptor, MCP-1, and TNF soluble receptor I. The inhibition of endogenous Wnt5a demonstrated that an autocrine Wnt5a loop is a major regulator of the NF-κB pathway in melanoma. Taken together, these results indicate that Wnt5a activates the NF-κB pathway and has an immunomodulatory effect on melanoma through the secretion of cytokines and chemokines.

scholarly journals Comparison of the Inhibitory Activities of 5,6-Dihydroergosterol Glycoside α- and β-Anomers on Skin Inflammation

Molecules ◽  
2019 ◽  
Vol 24 (2) ◽  
pp. 371 ◽  
Author(s):  
Tae Kim ◽  
Young Cho ◽  
HoonGyu Park ◽  
Tae Lee ◽  
Hakwon Kim
Keyword(s):  
Protein Expression ◽  
Inflammatory Diseases ◽  
Skin Barrier ◽  
Skin Inflammation ◽  
New Drugs ◽  
Inflammatory Biomarker ◽  
Cytokines And Chemokines ◽  
Inflammatory Stimuli ◽  
Mrna And Protein Expression ◽  
Chronic Skin

Chronic skin inflammatory diseases, such as atopic dermatitis, are associated with a dysfunctional skin barrier due to an increase in various inflammatory stimuli, for instance inflammatory cytokines and chemokines. In particular, CCL17 and CCL22 expression is increased in patients with chronic skin inflammation. In this study, we synthesized several α- and β-anomers of dihydroergosterol (DHE)-glycosides and assessed their effects on CCL17 and CCL22 expression. We confirmed that the β-anomers of DHE-glycosides were superior to α-anomers of DHE-glycosides in inhibiting CCL17 and CCL22 mRNA and protein expression. In addition, we determined that DHE-glycoside β-anomers showed strong inhibitory activity towards pro-inflammatory cytokine mRNA and protein expression, including that of TNF-α, IL-6, and IL-1β- in stimulated HaCaT cells. These results imply that DHE-glycoside α- and β-anomers should be separated during synthesis of drugs for chronic skin inflammation. Our results also suggest that β-anomers of DHE-glycosides may play an important role as new drugs for chronic skin inflammation because of their ability to inhibit the skin inflammatory biomarker proteins CCL17 and CCL22.

scholarly journals Loss of Caveolin-1 Gene Expression Accelerates the Development of Dysplastic Mammary Lesions in Tumor-Prone Transgenic Mice

2003 ◽  
Vol 14 (3) ◽  
pp. 1027-1042 ◽  
Author(s):  
Terence M. Williams ◽  
Michelle W.-C. Cheung ◽  
David S. Park ◽  
Babak Razani ◽  
Alex W. Cohen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Tumor Suppressor ◽  
Transgenic Mice ◽  
Histological Grade ◽  
Dominant Negative ◽  
Breast Cancers ◽  
Suppressor Activity ◽  
Caveolin 1 ◽  
Dominant Negative Mutation

Caveolin-1 is the principal structural component of caveolae microdomains, which represent a subcompartment of the plasma membrane. Several independent lines of evidence support the notion that caveolin-1 functions as a suppressor of cell transformation. For example, the human CAV-1 gene maps to a suspected tumor suppressor locus (D7S522/7q31.1) that is frequently deleted in a number of carcinomas, including breast cancers. In addition, up to 16% of human breast cancers harbor a dominant-negative mutation, P132L, in the CAV-1 gene. Despite these genetic associations, the tumor suppressor role of caveolin-1 still remains controversial. To directly assess the in vivo transformation suppressor activity of the caveolin-1 gene, we interbred Cav-1 (−/−) null mice with tumor-prone transgenic mice (MMTV-PyMT) that normally develop multifocal dysplastic lesions throughout the entire mammary tree. Herein, we show that loss of caveolin-1 gene expression dramatically accelerates the development of these multifocal dysplastic mammary lesions. At 3 wk of age, loss of caveolin-1 resulted in an approximately twofold increase in the number of lesions (foci per gland; 3.3 ± 1.0 vs. 7.0 ± 1.2) and an approximately five- to sixfold increase in the total area occupied by these lesions. Similar results were obtained at 4 wk of age. However, complete loss of caveolin-1 was required to accelerate the appearance of these dysplastic mammary lesions, because Cav-1 (+/−) heterozygous mice did not show any increases in foci development. We also show that loss of caveolin-1 increases the extent and the histological grade of these mammary lesions and facilitates the development of papillary projections in the mammary ducts. Finally, we demonstrate that cyclin D1 expression levels are dramatically elevated in Cav-1 (−/−) null mammary lesions, consistent with the accelerated appearance and growth of these dysplastic foci. This is the first in vivo demonstration that caveolin-1 can function as a transformation suppressor gene.

scholarly journals c-Fos Mediates Angiotensin II-Induced Aldosterone Production and Protein Synthesis in Bovine Adrenal Glomerulosa Cells

Endocrinology ◽  
2008 ◽  
Vol 150 (3) ◽  
pp. 1294-1302 ◽  
Author(s):  
Jose Maria Rincon Garriz ◽  
Catherine Suarez ◽  
Alessandro M. Capponi
Keyword(s):  
Gene Expression ◽  
Protein Synthesis ◽  
Angiotensin Ii ◽  
Protein Expression ◽  
Dominant Negative ◽  
Early Gene ◽  
Star Protein ◽  
Aldosterone Production ◽  
Glomerulosa Cells ◽  
Star Gene

Angiotensin II (AngII), potassium ion, and ACTH are the main factors controlling aldosterone biosynthesis in adrenal glomerulosa cells. AP-1 response elements for the immediate early gene products, c-Fos and c-Jun, have been identified, among others, in the promoter of the steroidogenic acute regulatory (StAR) protein gene, whose expression is acutely regulated by activators of aldosterone production. In bovine glomerulosa cells, AngII treatment led to a rapid and transient increase in c-fos mRNA expression, c-Fos protein expression, and c-Fos phosphorylation. Inhibition of the ERK1/2 MAPK pathway abolished the effect of AngII on c-fos mRNA, protein, and phosphorylation. EMSA and chromatin immunoprecipitation experiments demonstrated that c-Fos binds with c-Jun to the proximal StAR promoter and that AngII treatment increases the amount of c-Fos bound to the promoter. Overexpression of a dominant-negative form of c-Fos with adenoviral vectors inhibited StAR mRNA and StAR protein expression as well as aldosterone biosynthesis in response to AngII. The dominant-negative c-Fos also prevented the increase in protein synthesis induced by AngII in glomerulosa cells, as assessed by [3H]leucine incorporation. These results indicate that AngII rapidly induces c-Fos expression and posttranslational modifications. Furthermore, a heterodimeric c-Fos/c-Jun complex binds to the proximal StAR promoter in glomerulosa cells, thus activating StAR gene expression and acute aldosterone biosynthesis. Finally, c-Fos also contributes to other functional responses to the hormone, such as protein synthesis. In adrenal glomerulosa cells, angiotensin II induces expression and phosphorylation of c-Fos, which mediates the peptide’s effects on StAR gene expression, aldosterone production and protein synthesis.

scholarly journals Plasma Membrane CFTR Regulates RANTES Expression via Its C-Terminal PDZ-Interacting Motif

2003 ◽  
Vol 23 (2) ◽  
pp. 594-606 ◽  
Author(s):  
Kim Estell ◽  
Gavin Braunstein ◽  
Torry Tucker ◽  
Karoly Varga ◽  
James F. Collawn ◽  
...  
Keyword(s):  
Gene Expression ◽  
Plasma Membrane ◽  
Chloride Channel ◽  
Chloride Channels ◽  
Inflammatory Diseases ◽  
Airway Epithelial Cells ◽  
Dominant Negative ◽  
Cystic Fibrosis Gene ◽  
Channel Activity ◽  
Signaling Complex

ABSTRACT Despite the identification of 1,000 mutations in the cystic fibrosis gene product CFTR, there remains discordance between CFTR genotype and lung disease phenotype. The study of CFTR, therefore, has expanded beyond its chloride channel activity into other possible functions, such as its role as a regulator of gene expression. Findings indicate that CFTR plays a role in the expression of RANTES in airway epithelia. RANTES is a chemokine that has been implicated in the regulation of mucosal immunity and the pathogenesis of airway inflammatory diseases. Results demonstrate that CFTR triggers RANTES expression via a mechanism that is independent of CFTR's chloride channel activity. Neither pharmacological inhibition of CFTR nor activation of alternative chloride channels, including hClC-2, modulated RANTES expression. Through the use of CFTR disease-associated and truncation mutants, experiments suggest that CFTR-mediated transcription factor activation and RANTES expression require (i) insertion of CFTR into the plasma membrane and (ii) an intact CFTR C-terminal PDZ-interacting domain. Expression of constructs encoding wild-type or dominant-negative forms of the PDZ-binding protein EBP50 suggests that EBP50 may be involved in CFTR-dependent RANTES expression. Together, these data suggest that CFTR modulates gene expression in airway epithelial cells while located in a macromolecular signaling complex at the plasma membrane.

scholarly journals A Novel Therapeutic Reagent, KA-1002 for Alleviating Lysophosphatidic Acid-Mediated Inflammation Related Gene Expression in Swine Macrophages

Animals ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 534
Author(s):  
Hyeon-Jeong Hwang ◽  
Tamina Park ◽  
Miok Kim ◽  
Hee-su Shin ◽  
Wooyeon Hwang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Lysophosphatidic Acid ◽  
Acute Inflammation ◽  
Inflammatory Diseases ◽  
Regulatory Effect ◽  
Related Gene ◽  
Industrial Environment ◽  
Transcriptional Regulatory ◽  
Inflammatory Molecule ◽  
Novel Therapeutic

Stresses and various infectious reagents caused multiple inflammatory diseases in swine in a livestock industrial environment. Therefore, there is a need for an effective therapeutic or preventive agent that could alleviate chronic and acute inflammation. We found that lysophosphatidic acid (LPA), a stress-induced potent endogenous inflammatory molecule, causes a broad range-regulation of inflammation related genes inflammation in swine macrophages. We further investigated the genome scaled transcriptional regulatory effect of a novel LPA-signaling antagonist, KA-1002 on swine macrophages, inducing the alleviated LPA-mediated inflammation related gene expression. Therefore, KA-1002 could potentially serve as a novel therapeutic or preventive agent to maintain physiologically healthy and balanced conditions of pigs.

ΔNp73 /TAp73 Expression Ratio Is Associated with Ara-C-Induced Apoptosis Resistance in Acute Myeloid Leukemia.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4148-4148
Author(s):  
Antonio Roberto L. Araújo ◽  
Rodrigo A. Panepucci ◽  
Rafael H. Jacomo ◽  
Guilherme A. Souza ◽  
Amélia G. Araújo ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Expression ◽  
De Novo ◽  
Gene Expression Pattern ◽  
Dominant Negative ◽  
Protein Isoforms ◽  
Leukemic Cells ◽  
Apoptosis Resistance ◽  
Expression Ratio

Abstract The p73, a homologue to the tumor suppressor gene p53, is involved in oncogenesis, although its specific role remains unclear. This gene has two distinct promoters which encode for two protein isoforms with opposite transcriptional activities: full-length TAp73 and the ΔNp73, the latter differing by the lack of the N-terminal transactivating domain and, has a dominant-negative function towards p53 and TAp73 activity. In the present study, we have investigated, by Real Time PCR, the gene expression pattern of TAp73 and ΔNp73 in 147 samples from patients with de novo AML and 22 samples of CD34+ hematopoietic progenitor cells isolated from bone marrow of healthy donors. We detected a significantly higher expression of TAp73 and ΔNp73 genes in AML samples compared to normal hematopoietic progenitors. AML samples harboring the chromosomal translocations: PML-RARa, AML1-ETO and CBFβ-MYH11 presented significantly higher levels of the TAp73 than those negative for these rearrangements. In contrast, samples without these gene rearrangements expressed higher levels of the truncated form ΔNp73. No difference was observed among patients with PML-RARa, AML1-ETO and CBFβ-MYH11. In order to determine if the ratio between the relative expression of TAp73 and ΔNp73 genes was associated with differential susceptibility to pro-apoptotic stimuli, we performed an in vitro cytotoxicity assay using Ara-C. Blasts from 20 AML patients were incubated for 24 hours in a medium containing 100μg-ml of Ara-C and evaluated for apoptosis status using Anexin V and Propidium Iodite. The leukemic cells with a higher ΔNp73 / TAp73 ratio were significantly more apoptosis-resistant (Pearson’s R2: −0.607). We also assessed the protein expression by flow cytometry using saturating amounts of specific antibodies for each isoform and analyzing the mean fluorescence channel. Estimated protein expression corroborated the gene expression results. Our results suggest an association between the presence of PML-RARα, AML1-ETO or CBFβ-MYH11 chromosomal translocation and a higher TAp73 gene expression compared to ΔNp73 isoforms expression in AML samples. Moreover, the higher resistence to apoptosis detected in blasts with higher ΔNp73 / TAp73 values suggest that this pathway may contribute to prognosis.

Mechanistic dissection of dominant AIRE mutations in mouse models reveals AIRE autoregulation

2021 ◽  
Vol 218 (11) ◽  
Author(s):  
Yael Goldfarb ◽  
Tal Givony ◽  
Noam Kadouri ◽  
Jan Dobeš ◽  
Cristina Peligero-Cruz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Expression ◽  
Mouse Models ◽  
Chromatin Accessibility ◽  
Dominant Negative ◽  
Docking Site ◽  
Central Tolerance ◽  
Recessive Mutations ◽  
Underlying Mechanisms ◽  
Autoregulatory Mechanism

The autoimmune regulator (AIRE) is essential for the establishment of central tolerance and prevention of autoimmunity. Interestingly, different AIRE mutations cause autoimmunity in either recessive or dominant-negative manners. Using engineered mouse models, we establish that some monoallelic mutants, including C311Y and C446G, cause breakdown of central tolerance. By using RNAseq, ATACseq, ChIPseq, and protein analyses, we dissect the underlying mechanisms for their dominancy. Specifically, we show that recessive mutations result in a lack of AIRE protein expression, while the dominant mutations in both PHD domains augment the expression of dysfunctional AIRE with altered capacity to bind chromatin and induce gene expression. Finally, we demonstrate that enhanced AIRE expression is partially due to increased chromatin accessibility of the AIRE proximal enhancer, which serves as a docking site for AIRE binding. Therefore, our data not only elucidate why some AIRE mutations are recessive while others dominant, but also identify an autoregulatory mechanism by which AIRE negatively modulates its own expression.

scholarly journals Training-Associated Alterations In Equine Respiratory Immunity – A Multi-Omics Comparative Approach.

2021 ◽  
Author(s):  
Anna Eleonora Karagianni ◽  
Dominic Kurian ◽  
Eugenio Cillán-Garcia ◽  
Samantha Eaton ◽  
Thomas M. Wishart ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Expression ◽  
Airway Inflammation ◽  
Cell Function ◽  
Immune Cell ◽  
Methodological Approach ◽  
Training Period ◽  
Comparative Approach ◽  
Thoroughbred Racehorses ◽  
Gene And Protein Expression

Abstract Background: Neutrophilic airway inflammation is highly prevalent in racehorses in training, with the term mild to moderate equine asthma (MMEA) being applied to the majority of such cases. Our proposed study is largely derived from the strong association between MMEA in racehorses and their entry into a race training program; this has led to our primary aim of measuring the effect of race training on pulmonary immune cell function. The objectives of this study are to characterise the effect of training on the local pulmonary immune system and quantify the magnitude of effect by defining (a) the gene expression of tracheal wash (TW) derived cells and (b) the protein expression of TW samples derived from Thoroughbred racehorses prior to (T0) and following commencement of race training (T1). Results: Tracheal wash samples were obtained at both time points (T0, T1) from the same Thoroughbred horses (n=16). Gene expression of TW derived cells, determined by RNAseq and analysed by DEseq2 detected 2,138 differentially expressed (DE) genes during the training period; 1,122 of these were upregulated. The proteome of TW samples was also evaluated and contained 260 DE proteins during the training period; 103 of these were upregulated and the rest downregulated. Gene and protein sets were enriched for biological processes related to acute phase response and oxidative stress, as well as to immune response and inflammation. Many genes, including ISG15, ISG20, IFI35, SOCS1 and TRIM21, were highly enriched for IFN signaling. Interestingly, pathway analysis also highlighted genes and proteins related to haemopoietic processes.Conclusions: This study demonstrated TW samples to represent a rich source of airway cells, protein and RNA to study airway immunity in the horse and highlighted the benefits of a multi-omics methodological approach to studying the dynamics of equine airway immunity. Intense training induced quantifiable alterations in both gene and protein expression of airway derived cells is consistent with deregulation of airway immunity and haemopoietic abnormalities. Respectively, these findings likely reflect the known associations between race training and both airway inflammation and bleeding, in particular offering further insight into the potential mechanisms which underpin training associated airway inflammation.

Sign in / Sign up

Export Citation Format

Share Document