22 results on '"Vuppusetty C"'
Search Results
2. Effects of L-leucine in 5q- syndrome and other RPS14-deficient erythroblasts
- Author
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Yip, B H, Pellagatti, A, Vuppusetty, C, Giagounidis, A, Germing, U, Lamikanra, A A, Roberts, D J, Fernandez-Mercado, M, McDonald, E-J, Killick, S, Wainscoat, J S, and Boultwood, J
- Published
- 2012
- Full Text
- View/download PDF
3. Decreased Serum Sirtuin-1 in COPD
- Author
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Yanagisawa, S, Papaioannou, AI, Papaporfyriou, A, Baker, JR, Vuppusetty, C, Loukides, S, Barnes, PJ, and Ito, K
- Subjects
Male ,Vital Capacity ,DMEM, Dulbecco's Modified Eagle Medium ,Dlco, diffusing capacity for carbon monoxide ,HRCT, high-resolution CT ,Pulmonary Disease, Chronic Obstructive ,GOLD, Global Initiative for Chronic Obstructive Lung Disease ,FBS, fetal bovine serum ,NAD, nicotinamide adenine dinucleotide ,Sirtuin 1 ,Forced Expiratory Volume ,s120S, serum SIRT1 ,COPD ,Humans ,sirtuin-1 ,IC, inspiratory capacity ,RPMI, Roswell Park Memorial Institute medium ,Cells, Cultured ,Aged ,SIRT1, silent information regulator 2 homolog 1 ,RV, residual volume ,TLC, total lung capacity ,ELISA, enzyme-linked immunosorbent assay ,Middle Aged ,MRC, Medical Research Council ,Original Research: COPD ,respiratory tract diseases ,emphysema ,6MWD, 6-min walking distance ,biomarker ,BODE, BMI, airflow obstruction, dyspnea, and exercise capacity ,KCO%, diffusing capacity of the lung per unit volume ,Female ,Aado2, alveolar-arterial oxygen difference ,serum ,Biomarkers - Abstract
Background The protein deacetylase sirtuin-1 (SIRT1) is an antiaging molecule that is decreased in the lung in patients with COPD. Recently, SIRT1 was reported to be detectable in serum, but serum SIRT1 (s120S) levels have not yet been reported in patients with COPD. Methods Serum SIRT1 protein of all samples was measured by Western blot, and the SIRT1 protein band densities were calculated and compared with clinical parameters. Results Several molecular sizes of SIRT1, including 120 kDa (actual size) and fragments (102 and 75 kDa) were quantified by Western blot. Among them, only the 120-kDa s120S was significantly decreased in patients with COPD compared with the control subjects without COPD (s120S ratio in healthy subjects = 0.90 ± 0.34 vs those with COPD = 0.68 ± .24; P = .014) and was positively correlated with airway obstruction (FEV1/FVC, r = 0.31; P = .020); its severity measured by FEV1 % predicted (r = 0.29; P = .029). s120S also showed a positive correlation with BMI (r = 0.36; P = .0077) and diffusing capacity of the lung per unit volume (the carbon monoxide transfer coefficient: KCO%) (r = 0.32; P = .025). It was also significantly decreased with increasing severity of lung emphysema (r = –0.40; P = .027) and with a clinical history of frequent COPD exacerbations (infrequent vs frequent, 0.76 ± 0.20 vs 0.56 ± 0.26; P = .027). SIRT1 was not detected in supernatant of A549 and primary epithelial cells in normal culture conditions. Conclusions s120S was decreased in the patients with COPD, potentially as reflected by the reduced SIRT1 within cells as a result of oxidative stress, and might be a potential biomarker for certain disease characteristics of COPD
- Published
- 2017
4. MicroRNA-570 is a novel regulator of cellular senescence and inflammaging
- Author
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Baker, JR, Vuppusetty, C, Colley, T, Hassibi, S, Fenwick, PS, Donnelly, LE, Ito, K, and Barnes, PJ
- Subjects
Inflammation ,Male ,Proto-Oncogene Proteins c-jun ,Research ,Middle Aged ,p38 Mitogen-Activated Protein Kinases ,epithelial cells ,Cell Line ,MicroRNAs ,Oxidative Stress ,Pulmonary Disease, Chronic Obstructive ,Sirtuin 1 ,Humans ,COPD ,Female ,cell cycle ,Cellular Senescence ,Aged ,Protein Binding ,Signal Transduction ,miRNA - Abstract
Diseases of accelerated aging often occur together (multimorbidity), and their prevalence is increasing, with high societal and health care costs. Chronic obstructive pulmonary disease (COPD) is one such condition, in which one half of patients exhibit ≥4 age-related diseases. Diseases of accelerated aging share common molecular pathways, which lead to the detrimental accumulation of senescent cells. These senescent cells no longer divide but release multiple inflammatory proteins, known as the senescence-associated secretory phenotype, which may perpetuate and speed disease. Here, we show that inhibiting miR-570-3p, which is increased in COPD cells, reverses cellular senescence by restoring the antiaging molecule sirtuin-1. MiR-570-3p is induced by oxidative stress in airway epithelial cells through p38 MAP kinase-c-Jun signaling and drives senescence by inhibiting sirtuin-1. Inhibition of elevated miR-570-3p in COPD small airway epithelial cells, using an antagomir, restores sirtuin-1 and suppresses markers of cellular senescence (p16INK4a, p21Waf1, and p27Kip1), thereby restoring cellular growth by allowing progression through the cell cycle. MiR-570-3p inhibition also suppresses the senescence-associated secretory phenotype (matrix metalloproteinases-2/9, C-X-C motif chemokine ligand 8, IL-1β, and IL-6). Collectively, these data suggest that inhibiting miR-570-3p rejuvenates cells via restoration of sirtuin-1, reducing many of the abnormalities associated with cellular senescence.—Baker, J. R., Vuppusetty, C., Colley, T., Hassibi, S., Fenwick, P. S., Donnelly, L. E., Ito, K., Barnes, P. J. MicroRNA-570 is a novel regulator of cellular senescence and inflammaging.
- Published
- 2018
5. The dynamic shuttling of SIRT1 between cytoplasm and nuclei in bronchial epithelial cells by single and repeated cigarette smoke exposure
- Author
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Yanagisawa, S, Baker, JR, Vuppusetty, C, Koga, T, Colley, T, Fenwick, P, Donnelly, LE, Barnes, PJ, Ito, K, Yildirim, AÖ, and Yildirim, AÖ
- Subjects
0301 basic medicine ,Cytoplasm ,Pulmonology ,lcsh:Medicine ,medicine.disease_cause ,Biochemistry ,Epithelium ,Antioxidants ,chemistry.chemical_compound ,Habits ,Sirtuin 1 ,Animal Cells ,Medicine and Health Sciences ,Smoking Habits ,Public and Occupational Health ,lcsh:Science ,Multidisciplinary ,biology ,Chemistry ,Reverse Transcriptase Polymerase Chain Reaction ,Smoking ,3. Good health ,Cell biology ,Enzymes ,Dismutases ,Cellular Types ,Anatomy ,Cellular Structures and Organelles ,Research Article ,Cell type ,Substance-Related Disorders ,General Science & Technology ,Chronic Obstructive Pulmonary Disease ,Blotting, Western ,SOD2 ,Bronchi ,Superoxide dismutase ,03 medical and health sciences ,Mental Health and Psychiatry ,MD Multidisciplinary ,medicine ,Humans ,Phosphatidylinositol ,PI3K/AKT/mTOR pathway ,Cell Nucleus ,Messenger RNA ,Behavior ,Dose-Response Relationship, Drug ,Superoxide Dismutase ,lcsh:R ,Biology and Life Sciences ,Proteins ,Smoking Related Disorders ,Epithelial Cells ,Cell Biology ,Oxidative Stress ,030104 developmental biology ,Biological Tissue ,biology.protein ,Enzymology ,lcsh:Q ,Oxidative stress - Abstract
SIRT1 (silent information regulator 2 homolog 1) is a crucial cellular survival protein especially in oxidative stress environments, and has been thought to locate within the nuclei, but also known to shuttle between cytoplasm and nuclei in some cell types. Here, we show for the first time the dynamics of SIRT1 in the presence of single or concurrent cigarette smoke extract (CSE) exposure in human bronchial epithelial cells (HBEC). In BEAS-2B HBEC or primary HBEC, SIRT1 was localized predominantly in cytoplasm, and the CSE (3%) induced nuclear translocation of SIRT1 from cytoplasm in the presence of L-buthionine sulfoximine (an irreversible inhibitor of γ-glutamylcystein synthetase), mainly through the activation of phosphatidylinositol 3-kinase (PI3K) α subunit. This SIRT1 nuclear shuttling was associated with FOXO3a nuclear translocation and the strong induction of several anti-oxidant genes including superoxide dismutase (SOD) 2 and 3; therefore seemed to be an adaptive response. When BEAS-2B cells were pretreated with repeated exposure to a lower concentration of CSE (0.3%), the CSE-induced SIRT1 shuttling and resultant SOD2/3 mRNA induction were significantly impaired. Thus, this result offers a useful cell model to mimic the impaired anti-oxidant capacity in cigarette smoking-associated lung disease such as chronic obstructive pulmonary disease.
- Published
- 2018
6. Decreased phosphatase PTEN amplifies PI3K signaling and enhances pro-inflammatory cytokine release in COPD
- Author
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Yanagisawa, S, Baker, JR, Vuppusetty, C, Fenwick, P, Donnelly, LE, Ito, K, Barnes, PJ, and Wellcome Trust
- Subjects
Male ,PTEN ,REDOX REGULATION ,Physiology ,Respiratory System ,HUMAN LUNG CANCERS ,OBSTRUCTIVE PULMONARY-DISEASE ,PI3K ,TENSIN HOMOLOG ,chronic obstructive pulmonary disease ,TUMOR-SUPPRESSOR PTEN ,NEGATIVE REGULATION ,Cell Line ,Phosphatidylinositol 3-Kinases ,Pulmonary Disease, Chronic Obstructive ,Smoke ,GLUTATHIONE ,oxidative stress ,Humans ,COPD ,Phosphorylation ,Lung ,INDUCED SPUTUM ,Aged ,Inflammation ,IL-6 ,Science & Technology ,Smoking ,phosphatase and tensin homolog deleted from chromosome 10 ,PTEN Phosphohydrolase ,EPITHELIAL-CELLS ,Epithelial Cells ,0606 Physiology ,respiratory tract diseases ,1116 Medical Physiology ,PI3K IL-6 ,Cytokines ,INACTIVATION ,Female ,Life Sciences & Biomedicine ,Proto-Oncogene Proteins c-akt ,Research Article ,Signal Transduction - Abstract
The phosphatidylinositol 3-kinase (PI3K) pathway is activated in chronic obstructive pulmonary disease (COPD), but the regulatory mechanisms for this pathway are yet to be elucidated. Our aim was to determine the expression and role of phosphatase and tensin homolog deleted from chromosome 10 (PTEN), a negative regulator of the PI3K pathway, in COPD. PTEN expression and activity were measured in the peripheral lung of COPD patients compared to smoking and non-smoking controls. The direct influence of cigarette smoke extract (CSE) on PTEN expression was assessed using primary lung epithelial cells and a cell line (BEAS-2B) in the presence or absence of L-buthionine-sulfoximine (BSO) to deplete intracellular glutathione. The impact of PTEN knock-down by RNA interference on cytokine production was also examined. In peripheral lung, PTEN protein was significantly decreased in patients with COPD compared to the subjects without COPD (p < 0.001), and positively correlated with the severity of air-flow obstruction (FEV1 % predicted; r = 0.50; p = 0.0012), although no difference was observed in PTEN activity. Conversely, phosphorylated Akt, as a marker of PI3K activation, showed a negative correlation with PTEN protein levels (r = -0.41; p = 0.0042). Both in primary bronchial epithelial cells and BEAS-2B cell line, CSE decreased PTEN protein, which was reversed by N-acetylcysteine treatment. PTEN knock-down potentiated Akt phosphorylation and enhanced production of pro-inflammatory cytokines, such as IL-6, CXCL8, CCL2 and CCL5. In conclusion, oxidative stress reduces PTEN protein levels, which may result in increased PI3K signaling and amplification of inflammation in COPD.
- Published
- 2017
7. Decreased serum sirtuin-1 in chronic obstructive pulmonary disease
- Author
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Yanagisawa, S, Papaioannou, A, Papaporfyriou, A, Baker, J, Vuppusetty, C, Loukides, S, Barnes, PJ, Ito, K, and Wellcome Trust
- Subjects
Male ,PROTEIN DEACETYLASE ,Respiratory System ,Vital Capacity ,METABOLISM ,OBSTRUCTIVE PULMONARY-DISEASE ,Pulmonary Disease, Chronic Obstructive ,SIRT1 ,Critical Care Medicine ,INFLAMMATION ,Sirtuin 1 ,General & Internal Medicine ,Forced Expiratory Volume ,COPD ,Humans ,sirtuin-1 ,OXIDATIVE STRESS ,Cells, Cultured ,Aged ,Science & Technology ,1103 Clinical Sciences ,Middle Aged ,respiratory tract diseases ,emphysema ,CELLS ,biomarker ,Female ,DEPENDENT HISTONE DEACETYLASE ,AGING PROCESS ,Life Sciences & Biomedicine ,serum ,LUNG ,Biomarkers - Abstract
Background: The protein deacetylase sirtuin-1 (SIRT1) is an anti-aging molecule that is decreased in the lung from patients with chronic obstructive pulmonary disease (COPD). Recently, SIRT1 was reported to be detectable in serum, but serum SIRT1 levels have not yet been reported in patients with COPD. Methods: Serum SIRT1 was measured by Western blotting, and relative ratio of band density in samples against that of a positive control were calculated. Results: Several molecular sizes of SIRT1, including 120kDa (actual size) and fragments (102, 75kDa) were quantified by Western blotting. Among them, only the 120kDa serum SIRT1 (s120S) was significantly decreased in the patients with COPD compared to the control subjects without COPD (s120S ratio in healthy: 0.90±0.34, vs COPD: 0.68±0.24; p=0.014), and was positively correlated with airway obstruction (FEV1/ FVC; r=0.31; p=0.020) and its severity measured by FEV1 % predicted (r=0.29; p=0.029). Serum s120S also showed a positive correlation with body mass index (BMI; r=0.36; p=0.0077) and diffusing capacity of the lung per unit volume (KCO%; r=0.32; p=0.025). It was also significantly decreased with increasing severity of lung emphysema (r=-0.40, p=0.027) and with a clinical history of frequent COPD exacerbations (infrequent: 0.76±0.20 vs frequent: 0.56±0.26; p=0.027). SIRT1 was not detected in supernatant of A549 and primary epithelial cells in normal culture condition. Conclusions: Serum SIRT1 (s120S) was decreased in the patients with COPD, potentially as reflected by the reduced SIRT1 within cells as a result of oxidative stress, and might be a potential biomarkers for certain disease characteristics of COPD.
- Published
- 2017
8. Decreased Serum Sirtuin-1 in COPD
- Author
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Yanagisawa, S. Papaioannou, A.I. Papaporfyriou, A. Baker, J.R. Vuppusetty, C. Loukides, S. Barnes, P.J. Ito, K.
- Subjects
respiratory tract diseases - Abstract
Background The protein deacetylase sirtuin-1 (SIRT1) is an antiaging molecule that is decreased in the lung in patients with COPD. Recently, SIRT1 was reported to be detectable in serum, but serum SIRT1 (s120S) levels have not yet been reported in patients with COPD. Methods Serum SIRT1 protein of all samples was measured by Western blot, and the SIRT1 protein band densities were calculated and compared with clinical parameters. Results Several molecular sizes of SIRT1, including 120 kDa (actual size) and fragments (102 and 75 kDa) were quantified by Western blot. Among them, only the 120-kDa s120S was significantly decreased in patients with COPD compared with the control subjects without COPD (s120S ratio in healthy subjects = 0.90 ± 0.34 vs those with COPD = 0.68 ± 0.24; P =.014) and was positively correlated with airway obstruction (FEV1/FVC, r = 0.31; P =.020); its severity measured by FEV1 % predicted (r = 0.29; P =.029). s120S also showed a positive correlation with BMI (r = 0.36; P =.0077) and diffusing capacity of the lung per unit volume (the carbon monoxide transfer coefficient: KCO%) (r = 0.32; P =.025). It was also significantly decreased with increasing severity of lung emphysema (r = –0.40; P =.027) and with a clinical history of frequent COPD exacerbations (infrequent vs frequent, 0.76 ± 0.20 vs 0.56 ± 0.26; P =.027). SIRT1 was not detected in supernatant of A549 and primary epithelial cells in normal culture conditions. Conclusions s120S was decreased in the patients with COPD, potentially as reflected by the reduced SIRT1 within cells as a result of oxidative stress, and might be a potential biomarker for certain disease characteristics of COPD. © 2017 The Authors
- Published
- 2017
9. Developing chemical probes for the BET bromodomains
- Author
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Hewings, DS, Wang, M, Philpott, M, Fedorov, O, Uttarkar, S, Filippakopoulos, P, Picaud, S, Vuppusetty, C, Marsden, B, Heightman, TD, Knapp, S, Brennan, P, and Conway, SJ
- Published
- 2016
10. Oxidative stress dependent microRNA-34a activation via PI3Kα reduces the expression of sirtuin-1 and sirtuin-6 in epithelial cells
- Author
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Baker, JR, Vuppusetty, C, Colley, T, Papaioannou, AI, Fenwick, P, Donnelly, L, Ito, K, Barnes, PJ, and Wellcome Trust
- Subjects
PTEN Phosphohydrolase ,Sputum ,Antagomirs ,Down-Regulation ,Nuclear Proteins ,RNA-Binding Proteins ,Epithelial Cells ,Hydrogen Peroxide ,Article ,Cell Line ,Up-Regulation ,DNA-Binding Proteins ,MicroRNAs ,Oxidative Stress ,Phosphatidylinositol 3-Kinases ,Pulmonary Disease, Chronic Obstructive ,Sirtuin 1 ,Humans ,Sirtuins ,RNA, Messenger ,3' Untranslated Regions ,Lung ,Cellular Senescence - Abstract
Sirtuin-1 (SIRT1) and SIRT6, NAD(+)-dependent Class III protein deacetylases, are putative anti-aging enzymes, down-regulated in patients with chronic obstructive pulmonary disease (COPD), which is characterized by the accelerated ageing of the lung and associated with increased oxidative stress. Here, we show that oxidative stress (hydrogen peroxide) selectively elevates microRNA-34a (miR-34a) but not the related miR-34b/c, with concomitant reduction of SIRT1/-6 in bronchial epithelial cells (BEAS2B), which was also observed in peripheral lung samples from patients with COPD. Over-expression of a miR-34a mimic caused a significant reduction in both mRNA and protein of SIRT1/-6, whereas inhibition of miR-34a (antagomir) increased these sirtuins. Induction of miR-34a expression with H2O2 was phosphoinositide-3-kinase (PI3K) dependent as it was associated with PI3Kα activation as well as phosphatase and tensin homolog (PTEN) reduction. Importantly, miR-34a antagomirs increased SIRT1/-6 mRNA levels, whilst decreasing markers of cellular senescence in airway epithelial cells from COPD patients, suggesting that this process is reversible. Other sirtuin isoforms were not affected by miR-34a. Our data indicate that miR-34a is induced by oxidative stress via PI3K signaling, and orchestrates ageing responses under oxidative stress, therefore highlighting miR-34a as a new therapeutic target and biomarker in COPD and other oxidative stress-driven aging diseases.
- Published
- 2016
11. 3,5-Dimethylisoxazoles Act As Acetyl-lysine-mimetic Bromodomain Ligands
- Author
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Hewings, D, Wang, M, Philpott, M, Fedorov, O, Uttarkar, S, Filippakopoulos, P, Picaud, S, Vuppusetty, C, Marsden, B, Knapp, S, Conway, S, Heightman, T, and Society, American Chemical
- Subjects
Models, Molecular ,BRD4 ,Stereochemistry ,Chemistry & allied sciences ,Cell Cycle Proteins ,Protein Serine-Threonine Kinases ,Medical sciences ,Crystallography, X-Ray ,Ligands ,01 natural sciences ,Article ,Histones ,Structure-Activity Relationship ,03 medical and health sciences ,chemistry.chemical_compound ,Drug Discovery ,Humans ,Structure–activity relationship ,CREB-binding protein ,030304 developmental biology ,0303 health sciences ,Molecular Structure ,biology ,Cytotoxins ,010405 organic chemistry ,Chemistry ,Lysine ,Molecular Mimicry ,Nuclear Proteins ,Stereoisomerism ,Isoxazoles ,Phenylethyl Alcohol ,CREB-Binding Protein ,Protein Structure, Tertiary ,0104 chemical sciences ,3. Good health ,Chromatin ,Bromodomain ,Histone ,Biochemistry ,Acetylation ,biology.protein ,Molecular Medicine ,Bioisostere ,HeLa Cells ,Protein Binding ,Transcription Factors - Abstract
Histone-lysine acetylation is a vital chromatin post-translational modification involved in the epigenetic regulation of gene transcription. Bromodomains bind acetylated lysines, acting as readers of the histone-acetylation code. Competitive inhibitors of this interaction have antiproliferative and anti-inflammatory properties. With 57 distinct bromodomains known, the discovery of subtype-selective inhibitors of the histone-bromodomain interaction is of great importance. We have identified the 3,5 dimethylisoxazole moiety as a novel acetyl-lysine bioisostere, which displaces acetylated histone-mimicking peptides from bromodomains. Using X-ray crystallographic analysis, we have determined the interactions responsible for the activity and selectivity of 4-substituted 3,5-dimethylisoxazoles against a selection of phylogenetically diverse bromodomains. By exploiting these interactions, we have developed compound 4d, which has IC50 values of
- Published
- 2011
12. 3,5-Dimethylisoxazoles inhibit the bromodomain-histone protein-protein interaction
- Author
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Hewings, D, Wang, M, Philpott, M, Fedorov, O, Uttarkar, S, Filippakopoulos, P, Picaud, S, Vuppusetty, C, Marsden, B, Heightman, T, Knapp, S, Brennan, P, and Conway, S
- Published
- 2012
13. Multi-Modal Characterization of Monocytes in Idiopathic Pulmonary Fibrosis Reveals a Primed Type I Interferon Immune Phenotype.
- Author
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Fraser E, Denney L, Antanaviciute A, Blirando K, Vuppusetty C, Zheng Y, Repapi E, Iotchkova V, Taylor S, Ashley N, St Noble V, Benamore R, Hoyles R, Clelland C, Rastrick JMD, Hardman CS, Alham NK, Rigby RE, Simmons A, Rehwinkel J, and Ho LP
- Subjects
- Case-Control Studies, Cells, Cultured, Chemokine CCL2 blood, Flow Cytometry, Gene Expression Profiling, Humans, Idiopathic Pulmonary Fibrosis genetics, Idiopathic Pulmonary Fibrosis metabolism, Idiopathic Pulmonary Fibrosis pathology, Immunophenotyping, Interferon Type I genetics, Interleukin-6 blood, Lung metabolism, Lung pathology, Macrophage Colony-Stimulating Factor blood, Macrophages immunology, Macrophages metabolism, Monocytes metabolism, Phenotype, Receptors, IgG genetics, Receptors, IgG metabolism, Single-Cell Analysis, Idiopathic Pulmonary Fibrosis immunology, Interferon Type I metabolism, Lung immunology, Monocytes immunology
- Abstract
Idiopathic pulmonary fibrosis (IPF) is the most severe form of chronic lung fibrosis. Circulating monocytes have been implicated in immune pathology in IPF but their phenotype is unknown. In this work, we determined the immune phenotype of monocytes in IPF using multi-colour flow cytometry, RNA sequencing and corresponding serum factors, and mapped the main findings to amount of lung fibrosis and single cell transcriptomic landscape of myeloid cells in IPF lungs. We show that monocytes from IPF patients displayed increased expression of CD64 (FcγR1) which correlated with amount of lung fibrosis, and an amplified type I IFN response ex vivo . These were accompanied by markedly raised CSF-1 levels, IL-6, and CCL-2 in serum of IPF patients. Interrogation of single cell transcriptomic data from human IPF lungs revealed increased proportion of CD64
hi monocytes and "transitional macrophages" with higher expression of CCL-2 and type I IFN genes. Our study shows that monocytes in IPF patients are phenotypically distinct from age-matched controls, with a primed type I IFN pathway that may contribute to driving chronic inflammation and fibrosis. These findings strengthen the potential role of monocytes in the pathogenesis of IPF., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Fraser, Denney, Antanaviciute, Blirando, Vuppusetty, Zheng, Repapi, Iotchkova, Taylor, Ashley, St Noble, Benamore, Hoyles, Clelland, Rastrick, Hardman, Alham, Rigby, Simmons, Rehwinkel and Ho.)- Published
- 2021
- Full Text
- View/download PDF
14. MicroRNA-570 is a novel regulator of cellular senescence and inflammaging.
- Author
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Baker JR, Vuppusetty C, Colley T, Hassibi S, Fenwick PS, Donnelly LE, Ito K, and Barnes PJ
- Subjects
- Aged, Cell Line, Female, Humans, Male, MicroRNAs metabolism, Middle Aged, Oxidative Stress, Protein Binding, Proto-Oncogene Proteins c-jun metabolism, Pulmonary Disease, Chronic Obstructive physiopathology, Signal Transduction, Sirtuin 1 metabolism, p38 Mitogen-Activated Protein Kinases metabolism, Cellular Senescence physiology, Inflammation physiopathology, MicroRNAs physiology
- Abstract
Diseases of accelerated aging often occur together (multimorbidity), and their prevalence is increasing, with high societal and health care costs. Chronic obstructive pulmonary disease (COPD) is one such condition, in which one half of patients exhibit ≥4 age-related diseases. Diseases of accelerated aging share common molecular pathways, which lead to the detrimental accumulation of senescent cells. These senescent cells no longer divide but release multiple inflammatory proteins, known as the senescence-associated secretory phenotype, which may perpetuate and speed disease. Here, we show that inhibiting miR-570-3p, which is increased in COPD cells, reverses cellular senescence by restoring the antiaging molecule sirtuin-1. MiR-570-3p is induced by oxidative stress in airway epithelial cells through p38 MAP kinase-c-Jun signaling and drives senescence by inhibiting sirtuin-1. Inhibition of elevated miR-570-3p in COPD small airway epithelial cells, using an antagomir, restores sirtuin-1 and suppresses markers of cellular senescence (p16
INK4a , p21Waf1 , and p27Kip1 ), thereby restoring cellular growth by allowing progression through the cell cycle. MiR-570-3p inhibition also suppresses the senescence-associated secretory phenotype (matrix metalloproteinases-2/9, C-X-C motif chemokine ligand 8, IL-1β, and IL-6). Collectively, these data suggest that inhibiting miR-570-3p rejuvenates cells via restoration of sirtuin-1, reducing many of the abnormalities associated with cellular senescence.-Baker, J. R., Vuppusetty, C., Colley, T., Hassibi, S., Fenwick, P. S., Donnelly, L. E., Ito, K., Barnes, P. J. MicroRNA-570 is a novel regulator of cellular senescence and inflammaging.- Published
- 2019
- Full Text
- View/download PDF
15. The dynamic shuttling of SIRT1 between cytoplasm and nuclei in bronchial epithelial cells by single and repeated cigarette smoke exposure.
- Author
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Yanagisawa S, Baker JR, Vuppusetty C, Koga T, Colley T, Fenwick P, Donnelly LE, Barnes PJ, and Ito K
- Subjects
- Blotting, Western, Bronchi drug effects, Bronchi metabolism, Cell Nucleus metabolism, Cytoplasm metabolism, Dose-Response Relationship, Drug, Epithelial Cells metabolism, Humans, Oxidative Stress drug effects, Reverse Transcriptase Polymerase Chain Reaction, Bronchi cytology, Epithelial Cells drug effects, Sirtuin 1 metabolism, Smoking adverse effects
- Abstract
SIRT1 (silent information regulator 2 homolog 1) is a crucial cellular survival protein especially in oxidative stress environments, and has been thought to locate within the nuclei, but also known to shuttle between cytoplasm and nuclei in some cell types. Here, we show for the first time the dynamics of SIRT1 in the presence of single or concurrent cigarette smoke extract (CSE) exposure in human bronchial epithelial cells (HBEC). In BEAS-2B HBEC or primary HBEC, SIRT1 was localized predominantly in cytoplasm, and the CSE (3%) induced nuclear translocation of SIRT1 from cytoplasm in the presence of L-buthionine sulfoximine (an irreversible inhibitor of γ-glutamylcystein synthetase), mainly through the activation of phosphatidylinositol 3-kinase (PI3K) α subunit. This SIRT1 nuclear shuttling was associated with FOXO3a nuclear translocation and the strong induction of several anti-oxidant genes including superoxide dismutase (SOD) 2 and 3; therefore seemed to be an adaptive response. When BEAS-2B cells were pretreated with repeated exposure to a lower concentration of CSE (0.3%), the CSE-induced SIRT1 shuttling and resultant SOD2/3 mRNA induction were significantly impaired. Thus, this result offers a useful cell model to mimic the impaired anti-oxidant capacity in cigarette smoking-associated lung disease such as chronic obstructive pulmonary disease.
- Published
- 2018
- Full Text
- View/download PDF
16. Quercetin restores corticosteroid sensitivity in cells from patients with chronic obstructive pulmonary disease.
- Author
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Mitani A, Azam A, Vuppusetty C, Ito K, Mercado N, and Barnes PJ
- Subjects
- Adult, Aged, Case-Control Studies, Female, Humans, Leukocytes, Mononuclear drug effects, Leukocytes, Mononuclear pathology, Male, Middle Aged, Pulmonary Disease, Chronic Obstructive pathology, Quercetin therapeutic use, Smoke adverse effects, Tobacco Products adverse effects, Tumor Cells, Cultured, U937 Cells, Adrenal Cortex Hormones pharmacology, Pulmonary Disease, Chronic Obstructive drug therapy, Quercetin pharmacology
- Abstract
Corticosteroid resistance is a major barrier to the effective treatment of chronic obstructive pulmonary disease (COPD). Oxidative stress from cigarette smoke and chronic inflammation is likely to induce this corticosteroid insensitivity. Quercetin is a polyphenol that has been reported to be an active oxygen scavenger as well as a functional adenosine monophosphate-activated protein kinase (AMPK) activator. The aim of this study was to investigate the effect of quercetin on corticosteroid responsiveness in COPD cells. Corticosteroid sensitivity was examined in human monocytic U937 cells exposed to cigarette smoke extract (CSE) and peripheral blood mononuclear cells (PBMC) collected from patients with COPD. Corticosteroid sensitivity was determined as the dexamethasone concentration causing 40% inhibition of tumor necrosis factor alpha-induced CXCL8 production (Dex-IC
40 ) in the presence or absence of quercetin. In U937 cells, treatment with quercetin activated AMPK and induced expression of nuclear factor erythroid 2-related factor 2, and consequently reversed CSE-induced corticosteroid insensitivity. PBMC from patients with COPD showed corticosteroid insensitivity compared with those from healthy volunteers, and treatment with quercetin restored corticosteroid sensitivity. In conclusion, quercetin restores corticosteroid sensitivity, and has the potential to be a novel treatment in combination with corticosteroids in COPD.- Published
- 2017
- Full Text
- View/download PDF
17. Decreased phosphatase PTEN amplifies PI3K signaling and enhances proinflammatory cytokine release in COPD.
- Author
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Yanagisawa S, Baker JR, Vuppusetty C, Fenwick P, Donnelly LE, Ito K, and Barnes PJ
- Subjects
- Aged, Cell Line, Epithelial Cells metabolism, Female, Humans, Lung metabolism, Male, Oxidative Stress physiology, Phosphorylation physiology, Proto-Oncogene Proteins c-akt metabolism, Signal Transduction physiology, Smoke adverse effects, Smoking adverse effects, Cytokines metabolism, Inflammation metabolism, PTEN Phosphohydrolase metabolism, Phosphatidylinositol 3-Kinases metabolism, Pulmonary Disease, Chronic Obstructive metabolism
- Abstract
The phosphatidylinositol 3-kinase (PI3K) pathway is activated in chronic obstructive pulmonary disease (COPD), but the regulatory mechanisms for this pathway are yet to be elucidated. The aim of this study was to determine the expression and role of phosphatase and tensin homolog deleted from chromosome 10 (PTEN), a negative regulator of the PI3K pathway, in COPD. PTEN protein expression was measured in the peripheral lung of COPD patients compared with smoking and nonsmoking controls. The direct influence of cigarette smoke extract (CSE) on PTEN expression was assessed using primary lung epithelial cells and a cell line (BEAS-2B) in the presence or absence of l-buthionine-sulfoximine (BSO) to deplete intracellular glutathione. The impact of PTEN knockdown by RNA interference on cytokine production was also examined. In peripheral lung, PTEN protein was significantly decreased in patients with COPD compared with the subjects without COPD ( P < 0.001) and positively correlated with the severity of airflow obstruction (forced expiratory volume in 1-s percent predicted; r = 0.50; P = 0.0012). Conversely, phosphorylated Akt, as a marker of PI3K activation, showed a negative correlation with PTEN protein levels ( r = -0.41; P = 0.0042). In both primary bronchial epithelial cells and BEAS-2B cells, CSE decreased PTEN protein, which was reversed by N -acetyl cysteine treatment. PTEN knockdown potentiated Akt phosphorylation and enhanced production of proinflammatory cytokines, such as IL-6, CXCL8, CCL2, and CCL5. In conclusion, oxidative stress reduces PTEN protein levels, which may result in increased PI3K signaling and amplification of inflammation in COPD., (Copyright © 2017 the American Physiological Society.)
- Published
- 2017
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18. Restoration of Corticosteroid Sensitivity in Chronic Obstructive Pulmonary Disease by Inhibition of Mammalian Target of Rapamycin.
- Author
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Mitani A, Ito K, Vuppusetty C, Barnes PJ, and Mercado N
- Subjects
- Adrenal Cortex Hormones therapeutic use, Aged, Drug Resistance immunology, Female, Histone Deacetylase 2 drug effects, Histone Deacetylase 2 physiology, Humans, Immunosuppressive Agents immunology, Immunosuppressive Agents pharmacology, Immunosuppressive Agents therapeutic use, Male, Middle Aged, Oxidative Stress physiology, Phosphatidylinositol 3-Kinases drug effects, Phosphatidylinositol 3-Kinases physiology, Proto-Oncogene Proteins c-akt drug effects, Proto-Oncogene Proteins c-akt physiology, Proto-Oncogene Proteins c-jun physiology, Pulmonary Disease, Chronic Obstructive immunology, Pulmonary Disease, Chronic Obstructive physiopathology, Sirolimus immunology, Sirolimus therapeutic use, Smoking adverse effects, Smoking physiopathology, TOR Serine-Threonine Kinases physiology, U937 Cells drug effects, p38 Mitogen-Activated Protein Kinases drug effects, p38 Mitogen-Activated Protein Kinases physiology, Adrenal Cortex Hormones pharmacology, Drug Resistance drug effects, Oxidative Stress drug effects, Proto-Oncogene Proteins c-jun drug effects, Pulmonary Disease, Chronic Obstructive drug therapy, Sirolimus pharmacology, TOR Serine-Threonine Kinases drug effects
- Abstract
Rationale: Corticosteroid resistance is a major barrier to the effective treatment of chronic obstructive pulmonary disease (COPD). Several molecular mechanisms have been proposed, such as activations of the phosphoinositide-3-kinase/Akt pathway and p38 mitogen-activated protein kinase. However, the mechanism for corticosteroid resistance is still not fully elucidated., Objectives: To investigate the role of mammalian target of rapamycin (mTOR) in corticosteroid sensitivity in COPD., Methods: The corticosteroid sensitivity of peripheral blood mononuclear cells collected from patients with COPD, smokers, and nonsmoking control subjects, or of human monocytic U937 cells exposed to cigarette smoke extract (CSE), was quantified as the dexamethasone concentration required to achieve 30% inhibition of tumor necrosis factor-α-induced CXCL8 production in the presence or absence of the mTOR inhibitor rapamycin. mTOR activity was determined as the phosphorylation of p70 S6 kinase, using Western blotting., Measurements and Main Results: mTOR activity was increased in peripheral blood mononuclear cells from patients with COPD, and treatment with rapamycin inhibited this as well as restoring corticosteroid sensitivity. In U937 cells, CSE stimulated mTOR activity and c-Jun expression, but pretreatment with rapamycin inhibited both and also reversed CSE-induced corticosteroid insensitivity., Conclusions: mTOR inhibition by rapamycin restores corticosteroid sensitivity via inhibition of c-Jun expression, and thus mTOR is a potential novel therapeutic target for COPD.
- Published
- 2016
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19. Activation of the mTOR pathway by the amino acid (L)-leucine in the 5q- syndrome and other ribosomopathies.
- Author
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Boultwood J, Yip BH, Vuppusetty C, Pellagatti A, and Wainscoat JS
- Subjects
- Anemia, Diamond-Blackfan drug therapy, Anemia, Diamond-Blackfan genetics, Anemia, Diamond-Blackfan pathology, Anemia, Macrocytic drug therapy, Anemia, Macrocytic genetics, Anemia, Macrocytic pathology, Animals, Chromosome Deletion, Chromosomes, Human, Pair 5 genetics, Chromosomes, Human, Pair 5 metabolism, Erythroblasts drug effects, Erythroblasts metabolism, Erythroblasts pathology, Gene Expression Regulation drug effects, Haploinsufficiency, Humans, Leucine metabolism, Protein Biosynthesis, Ribosomal Proteins genetics, Ribosomal Proteins metabolism, Ribosomes genetics, Ribosomes pathology, TOR Serine-Threonine Kinases genetics, TOR Serine-Threonine Kinases metabolism, Anemia, Diamond-Blackfan metabolism, Anemia, Macrocytic metabolism, Leucine pharmacology, Ribosomes drug effects, Signal Transduction drug effects
- Abstract
Patients with the 5q- syndrome and Diamond-Blackfan anemia (DBA) suffer from a severe macrocytic anemia. The 5q- syndrome and DBA are disorders of aberrant ribosome biogenesis (ribosomopathies) and haploinsufficiency of the ribosomal protein genes RPS14 and RPS19, respectively, underlies the anemia found in these disorders. Erythroblasts obtained from patients with the 5q- syndrome and DBA show impaired mRNA translation and this defect in translation may represent a potential therapeutic target in these ribosomopathies. There are some indications that the amino acid l-leucine, a translation enhancer, may have some efficacy in this group of disorders. Recent studies have shown that l-leucine treatment of zebrafish and murine models of the 5q- syndrome and DBA results in a marked improvement in the anemia. l-leucine treatment of RPS14-deficient and RPS19-deficient erythroblasts and erythroblasts from patients with the 5q- syndrome has been shown to result in an increase in cell proliferation, erythroid differentiation and mRNA translation in culture. l-leucine has been shown to improve hemoglobin levels and transfusion independence in a patient with DBA. l-leucine activates the mTOR (mammalian target of rapamycin) signaling pathway that controls cell growth and mRNA translation. There is evidence to suggest that the promotion of translation via the mTOR pathway by l-leucine is the mechanism that underlies the enhanced erythroid progenitor cell growth and differentiation observed in animal and cellular models of the 5q- syndrome and DBA treated with this amino acid. These data support the rationale for clinical trials of l-leucine as a therapeutic agent for the 5q- syndrome and DBA., (Copyright © 2012 Elsevier Ltd. All rights reserved.)
- Published
- 2013
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20. A protein deacetylase SIRT1 is a negative regulator of metalloproteinase-9.
- Author
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Nakamaru Y, Vuppusetty C, Wada H, Milne JC, Ito M, Rossios C, Elliot M, Hogg J, Kharitonov S, Goto H, Bemis JE, Elliott P, Barnes PJ, and Ito K
- Subjects
- Animals, Cell Line, Gene Expression Regulation, Humans, Hydrogen Peroxide, Inflammation, Lung pathology, Macrophages, Matrix Metalloproteinase 9 analysis, Matrix Metalloproteinase 9 genetics, Mice, Monocytes, Oxidative Stress, Promoter Regions, Genetic, Pulmonary Disease, Chronic Obstructive metabolism, Pulmonary Disease, Chronic Obstructive pathology, Sirtuin 1, Sirtuins analysis, Sirtuins genetics, Smoke adverse effects, Nicotiana, Matrix Metalloproteinase 9 metabolism, Sirtuins physiology
- Abstract
Inappropriate elevation of matrix metalloproteinase-9 (MMP9) is reported to be involved in the pathogenesis of chronic obstructive pulmonary disease (COPD). The object of this study was to identify the molecular mechanism underlying this increase of MMP9 expression, and here we show that oxidative stress-dependent reduction of a protein deacetylase, SIRT1, known as a putative antiaging enzyme, causes elevation of MMP9 expression. A sirtuin inhibitor, splitomycin, and SIRT1 knockdown by RNA interference led an increase in MMP9 expression in human monocytic U937 cells and in primary sputum macrophages, which was detected by RT-PCR, Western blot, activity assay, and zymography. In fact, the SIRT1 level was significantly decreased in peripheral lungs of patients with COPD, and this increase was inversely correlated with MMP9 expression and MMP9 promoter activation detected by a chromatin immunoprecipitation assay. H(2)O(2) reduced SIRT1 expression and activity in U937 cells; furthermore, cigarette smoke exposure also caused reduction of SIRT1 expression in lung tissue of A/J mice, with concomitant elevation of MMP9. Intranasal treatment of a selective and novel SIRT1 small molecule activator, SRT2172, blocked the increase of MMP9 expression in the lung as well as pulmonary neutrophilia and the reduction in exercise tolerance. Thus, SIRT1 is a negative regulator of MMP9 expression, and SIRT1 activation is implicated as a novel therapeutic approach to treating chronic inflammatory diseases, in which MMP9 is abundant.
- Published
- 2009
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21. Nitration of distinct tyrosine residues causes inactivation of histone deacetylase 2.
- Author
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Osoata GO, Yamamura S, Ito M, Vuppusetty C, Adcock IM, Barnes PJ, and Ito K
- Subjects
- Cell Line, Histone Deacetylase 2, Histone Deacetylases genetics, Humans, Oxidative Stress, Proteasome Endopeptidase Complex metabolism, Repressor Proteins genetics, Tyrosine genetics, Epigenesis, Genetic, Histone Deacetylases metabolism, Nitrates metabolism, Repressor Proteins metabolism, Tyrosine metabolism
- Abstract
Histone deacetylases (HDACs) are key molecules involved in epigenetic regulation of gene expression. We have previously demonstrated that oxidative stress caused a reduction in HDAC2, resulting in amplified inflammation and reduced corticosteroid responsiveness. Here we showed nitrative/oxidative stress reduced HDAC2 expression via nitration of distinct tyrosine residues. Peroxynitrite, hydrogen peroxide and cigarette smoke-conditioned medium reduced HDAC2 expression in A549 epithelial cells in vitro. This reduction was due to increased proteasomal degradation following ubiquitination rather than reduction of mRNA expression or stability. HDAC2 was nitrated under nitrative/oxidative stress and in the peripheral lung tissues of smokers and patients with chronic obstructive pulmonary disease. Mutagenesis studies replacing tyrosine (Y) residues with alanine revealed that Y253 is at least partly responsible for the proteasomal degradation of HDAC2 under nitrative stress. Thus, nitration of distinct tyrosine residues modifies both the expression and activity of HDAC2, having an impact on epigenetic regulation.
- Published
- 2009
- Full Text
- View/download PDF
22. Steroid-resistant neutrophilic inflammation in a mouse model of an acute exacerbation of asthma.
- Author
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Ito K, Herbert C, Siegle JS, Vuppusetty C, Hansbro N, Thomas PS, Foster PS, Barnes PJ, and Kumar RK
- Subjects
- Acute Disease, Animals, Asthma genetics, Asthma metabolism, Biomarkers, Cell Nucleus metabolism, Disease Models, Animal, Female, Gene Expression Regulation, Inflammation genetics, Inflammation immunology, Inflammation metabolism, Inflammation pathology, Mice, Mice, Inbred BALB C, NF-kappa B metabolism, Oxidative Stress, RNA, Messenger genetics, Asthma immunology, Asthma pathology, Drug Resistance drug effects, Neutrophils drug effects, Neutrophils immunology, Steroids pharmacology
- Abstract
Neutrophilic inflammation in acute exacerbations of asthma tends to be resistant to treatment with glucocorticoids. This may be related to decreased activity and expression of histone deacetylase-2 (HDAC2), which down-regulates expression of proinflammatory genes via recruitment to the glucocorticoid receptor complex. We assessed airway inflammation and response to steroid treatment in a novel mouse model of an acute exacerbation of chronic asthma. Systemically sensitized mice received low-level challenge with aerosolized ovalbumin for 4 weeks, followed by a single moderate-level challenge to induce enhanced inflammation in distal airways. We assessed the effects of pre-treatment with dexamethasone on the accumulation of inflammatory cells in the airways, airway responsiveness to methacholine, expression and enzymatic activity of nuclear proteins including histone acetyl transferase (HAT) and HDAC2, and levels of transcripts for neutrophil chemoattractant and survival cytokines. Dexamethasone suppressed inflammation associated with eosinophil and T-lymphocyte recruitment, but did not prevent neutrophil accumulation or development of airway hyperresponsiveness. Increased activity of HAT was suppressed by steroid treatment, but the marked diminution of HDAC2 activity and increased activity of nuclear factor-kappaB were not reversed. Correspondingly, elevated expression of mRNA for TNF-alpha, granulocyte-macrophage colony-stimulating factor, IL-8, and p21(waf) were also not suppressed by dexamethasone. Levels of lipid peroxidation and protein nitration products were elevated in the acute exacerbation model. We conclude that impaired nuclear recruitment of HDAC2 could be an important mechanism of steroid resistance of the neutrophilic inflammation in exacerbations of asthma. Oxidative stress may contribute to decreased HDAC2 activity.
- Published
- 2008
- Full Text
- View/download PDF
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