Peter J Barnes

Brocklehurst's chapter provides an excellent overview of the understanding of asthma and its treatment in 1983, but much has changed in our understanding of asthma and there have been remarkable improvements in its management over the last 40 years. Mast cells are still considered to be a key effector cell through the release of bronchoconstrictor mediators, but several other cell types are involved including eosinophils and structural cells regulated by type 2 (T2) immunity and a network of cytokines. Blocking single mediators, such as leukotrienes or prostaglandins, is less effective than corticosteroids, which are effective by inhalation in almost all patients. The widespread use of inhaled corticosteroids, usually given in combination with a long-acting β2-agonist, controls asthma in most patients, with a marked reduction in exacerbations and mortality. For patients with severe eosinophilic asthma, biologics that …

Background: Extracellular vesicles (EVs) are important mediators of intercellular communication. In COPD, EVs spread cellular senescence by transferring selected microRNAs to neighbouring healthy cells. Thus, EVs participate in the accumulation of senescent cells in tissues which may account for disease progression. However, the mechanism of EV uptake by recipient cells is unknown.Aim: To compare the effect of endocytosis inhibitors on the internalisation of EVs in recipient epithelial cells.Methods: EVs were isolated from BEAS2B cell media by ultracentrifugation and stained with the lipophilic membrane dye, pkh67. Recipient BEAS2B cells were cultured with and without drugs that inhibit different mechanisms of endocytosis (pitstop, cytochalasin b, cytochalasin d, 30μM dynasore, nocodazole and β-cyclodextrin) and pkh67-labelled EVs for up to 12h. Internalisation of EVs in recipient cells was analysed by …

Tumor-necrosis factor (TNF) is recognized as a therapeutic target in inflammatory diseases, including asthma. In severe forms of asthma, biologics such as anti-TNF are rendered to be investigated as therapeutic options in severe asthma. Hence, this work is done to assess the efficacy and safety of anti-TNF as a supplementary therapy for patients with severe asthma. A systematic search of 3 databases (Cochrane Central Register of Controlled Trials, MEDLINE, ClinicalTrials.gov) was performed to identify for published and unpublished randomized controlled trials comparing anti-TNF (etanercept, adalimumab, infliximab, certolizumab pegol, golimumab) with placebo in patients diagnosed with persistent or severe asthma. Random-effects model was used to estimate risk ratios and mean differences (MDs) with confidence intervals (95% CIs). PROSPERO registration number is CRD42020172006. Four trials with 489 …

The inflammatory environment which protects against viral infections such as SARS-CoV-2 is dynamic. Elucidating the interaction of immune mediators will provide insight into disease pathogenesis and assist the development of therapeutics to treat disease caused by viral insults.Using previously published upper respiratory tract network analysis, we dissected clinical symptom severity (FLU-PRO questionnaire) and self-reported clinical recovery based on immune node clusters from 139 participants in a community-based randomised clinical trial in early onset SARS-CoV-2 (The STOIC study, NCT04416399). Primary outcome events reported included urgent care, emergency department visit or hospitalisation for COVID-19. Nodes at study enrolment were defined as interferon, innate immunity-like, chemokine-dominant, and mucosal immunity-like. Participants were stratified based on viral burden (30-cycle …

Optimizing asthma diagnosis is an essential part of global strategies to reduce the excessive illness burden from asthma. New understanding about how to address the complexity and heterogeneity of the different forms of asthma means that asthma diagnosis now requires a compound diagnostic approach and label. Eliciting the typical symptoms and abnormal physiology of variable airflow limitation permits the recognition of asthma, and the identification of further features, such as eosinophilic or type 2 inflammation, allows a compound diagnostic label of eosinophilic asthma. This conveys key information about future exacerbation risk and likely treatment responsiveness. Treatable traits are a useful way to implement this new approach to diagnosis. Targeted assessment is used to inform a specific treatment plan in a pragmatic and iterative process. (c) 2022 Published by Elsevier Inc. on behalf of the American …

The Global Initiative for Chronic Obstructive Lung Disease (GOLD) has published the complete 2023 GOLD report, which can be freely downloaded from its web page (www. goldcopd. org) together with a “pocket guide” and “teaching slide set”(1). It contains important changes compared to earlier versions, and incorporates 387 new references (1). Here, we present an executive summary of this GOLD 2023 report (1) that summarizes aspects that a) are relevant from a clinicians perspective and b) updates evidence published since the prior executive summary in 2017.

BackgroundIn 2008, a dedicated American Thoracic Society (ATS)/European Respiratory Society (ERS) task force published a paper on the possible use and limitations of clinical outcomes and biomarkers to evaluate the impact of pharmacological therapy in COPD patients. Since then our scientific understanding of COPD has increased considerably since then, there has been a progressive shift from a one-size-fits-all diagnostic/therapeutic approach to a personalized approach, and many new treatments currently in development will require new endpoints to evaluate their efficacy adequately.ObjectivesThe emergence of several new relevant outcome measures motivated the authors to review advances in the field and highlight the need to update the content of the original report.MethodsThe authors separately created search strategies for the literature, primarily based on their opinions and assessments supported by carefully chosen references. No centralized examination of the literature or uniform criteria for including or excluding evidence were used.ResultsEndpoints, outcomes and biomarkers have been revisited. The limitations of some of those reported in the ERS/ATS task force document have been highlighted. In addition, new tools that may be useful, especially in evaluating personalized therapy, have been described.ConclusionsSince the'label-free'treatable traits approach is becoming an important step toward precision medicine, future clinical trials should focus on highly prevalent treatable traits, which will influence the choice of outcomes and markers to be considered. The use of the new tools, particularly combination endpoints …

Increasing evidence suggests that there is acceleration of lung ageing in chronic lung diseases, such as chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF), with the accumulation of senescent cells in the lung. Senescent cells fail to repair tissue damage and release an array of inflammatory proteins, known as the senescence-associated secretory phenotype, which drive further senescence and disease progression. This suggests that targeting cellular senescence with senotherapies may treat the underlying disease process in COPD and IPF and thus reduce disease progression and mortality. Several existing or future drugs may inhibit the development of cellular senescence which is driven by chronic oxidative stress (senostatics), including inhibitors of PI3K-mTOR signalling pathways, antagomirs of critical microRNAs and novel antioxidants. Other drugs (senolytics) selectively …

In an attempt to address issues related to global warming contributed to by the use of pressurised, metered-dose inhalers (pMDIs), Wilkinson et al (1) have succeeded in generating a great deal of negative, potentially harmful media interest for patients who currently rely on these devices. They analysed the potential impact of switching therapy from pMDIs to dry powder inhalers (DPIs) in terms of both changes in greenhouse gas emissions and costs to the UK National Health Service based on prescribing information in England alone. This strategy was apparently devised in a vacuum, without regard to implications for threats to patient wellbeing and safety as a consequence of their being deprived of access to pMDI therapy for obstructive airways disease (asthma and COPD). Physicians are obliged to consider many factors when selecting the most appropriate inhaler device for patients other than the cost to the …

Exposure to air pollution is a major contributor to the pathogenesis of COPD worldwide. Indeed, most recent estimates suggest that 50% of the total attributable risk of COPD may be related to air pollution. In response, the Global Initiative for Chronic Obstructive Lung Disease (GOLD) Scientific Committee performed a comprehensive review on this topic, qualitatively synthesised the evidence to date and proffered recommendations to mitigate the risk. The review found that both gaseous and particulate components of air pollution are likely contributors to COPD. There are no absolutely safe levels of ambient air pollution and the relationship between air pollution levels and respiratory events is supra-linear. Wildfires and extreme weather events such as heat waves, which are becoming more common owing to climate change, are major threats to COPD patients and acutely increase their risk of morbidity and mortality …

Background COPD is associated with cellular senescence and fibrosis. Extracellular vesicles (EVs) are membrane-derived vesicles involved in intercellular communication. EVs contain miRNAs, mRNA and proteins and have been implicated in COPD to induce senescence and the transition of fibroblast to myofibroblasts. This study examined whether EVs derived from COPD fibroblasts drive senescence in healthy recipient fibroblasts. Changes in expression of p21CIP1 and alpha-smooth muscle actin (αSMA) were chosen as markers of senescence and transition of fibroblasts to myofibroblasts respectively.Methods Large EVs, and small EVs were isolated from media from non-smoker (NS) and COPD fibroblasts cultured with or without H2O2. EVs were labelled with phk67 and uptake measured by flow cytometry. Healthy recipient fibroblasts were cultured with EVs or EV-free media for 24h and 48h and protein …

IL-36 cytokines are members of the IL-1 superfamily and consist of three receptor agonists (IL-36a, IL-36b, and IL-36g) and two receptor antagonists (IL-36Ra and IL-38). These bind to the IL-36 receptor (IL-36R), forming a heterodimer with IL-1R accessory protein (IL-1RAcP). Formation of the heterodimer activates the adaptor protein MyD88, MAPK (mitogen-activated protein kinase), and NF-kB signaling pathways, inducing proinflammatory cytokines and antiviral interferon signatures (1, 2). Respiratory viral infections are a major cause of adverse events in those with chronic respiratory disease, especially in exacerbations of chronic obstructive pulmonary disease (COPD)(3). IL-1 and IL-33 have been extensively studied during viral infections and exacerbations. IL-1 levels are increased during COPD exacerbations, and IL-1 inhibition reduces lung inflammation in mice exposed to cigarette smoke and influenza …

Current treatments fail to modify the underlying pathophysiology and disease progression of chronic obstructive pulmonary disease (COPD), necessitating alternative therapies. Here, we show that COPD subjects have increased IL-36γ and decreased IL-36 receptor antagonist (IL-36Ra) in bronchoalveolar and nasal fluid compared with control subjects. IL-36γ is derived from small airway epithelial cells (SAEC) and is further induced by a viral mimetic, whereas IL-36Ra is derived from macrophages. IL-36γ stimulates release of the neutrophil chemoattractants CXCL1 and CXCL8, as well as elastolytic matrix metalloproteinases (MMPs) from small airway fibroblasts (SAF). Proteases released from COPD neutrophils cleave and activate IL-36γ, thereby perpetuating IL-36 inflammation. Transfer of culture media from SAEC to SAF stimulated release of CXCL1, which was inhibited by exogenous IL-36Ra. The use of a …

We presenta robust, long‐range optical autofocus system for microscopy utilizing machine learning. This can be useful for experiments with long image data acquisition times that may be impacted by defocusing resulting from drift of components, for example due to changes in temperature or mechanical drift. It is also useful for automated slide scanning or multiwell plate imaging where the sample(s) to be imaged may not be in the same horizontal plane throughout the image data acquisition. To address the impact of (thermal or mechanical) fluctuations over time in the optical autofocus system itself, we utilize a convolutional neural network (CNN) that is trained over multiple days to account for such fluctuations. To address the trade‐off between axial precision and range of the autofocus, we implement orthogonal optical readouts with separate CNN training data, thereby achieving an accuracy well within the 600 …

Cellular senescence, when cells stop dividing, is a key feature of cell aging and may play an important role in the progression of COPD. Senescent cells accumulate in the lungs of COPD patients and release a variety of inflammatory proteins, cause fibrosis of small airways, and defective tissue repair leading to emphysema. The molecular pathways that lead to senescence and a loss of anti-aging molecules, such as sirtuins, are now better understood. Senescence may spread via microRNAs in extracellular vesicles, resulting in comorbidities. Identification of senescence mechanisms may result in new therapies (senotherapies), including senostatics that inhibit these pathways and senolytics that remove senescent cells. Understanding the pathways that lead to cellular senescence in COPD lungs may lead to new treatments that may reduce disease progression, mortality and comorbidities.

RATIONALE Chronic obstructive pulmonary disease (COPD) is associated with accelerated lung aging and increased pulmonary cellular senescence. Senescent cells are thought to be cleared by macrophage phagocytosis, but the mechanism behind this is not fully understood. COPD macrophages show reduced phagocytosis of bacteria and apoptotic cells but whether clearance of senescent cells is also impaired is unknown. OBJECTIVES To establish a flow cytometric phagocytosis assay measuring clearance of senescent BEAS-2B cells by monocyte-derived macrophages (MDM) in vitro and to use this to investigate the clearance of senescent airway epithelial cells by MDM from COPD and age-matched non-smoker (NS) subjects. METHODS Senescent airway epithelial BEAS-2B cell populations (60% senescence/senescence associatedβ-galactosidase staining) were generated by repeated exposure to 50nM …

Background Chronic obstructive pulmonary disease (COPD) is an inflammatory lung condition, associated with accelerated lung aging. Immune cells in aging diseases display features of senescence (immunosenescence) and chronic inflammation (inflammaging). COPD macrophages are more pro-inflammatory and have defective phagocytic function, but features of senescence in COPD macrophages is not well studied.Aim To examine expression of senescence markers in macrophages from healthy and COPD subjects, in the presence and absence of oxidative stress (H2O2).Methods Peripheral blood monocytes were isolated from age-matched non-smokers (NS) (n=9) and COPD patients (n=12) and then cultured in GM-CSF for 12 days to generate monocyte-derived macrophages (MDM). MDM were then treated with ±200µM H2O2 for 48h and expression of senescence markers was measured using qPCR …

There is a marked increase in oxidative stress in the lungs of patients with COPD, as measured by increased exhaled 8-isoprostane, ethane, and hydrogen peroxide in the breath. The lung may be exposed to exogenous oxidative stress from cigarette smoking and indoor or outdoor air pollution and to endogenous oxidative stress from reactive oxygen species released from activated inflammatory cells, particularly neutrophils and macrophages, in the lungs. Oxidative stress in COPD may be amplified by a reduction in endogenous antioxidants and poor intake of dietary antioxidants. Oxidative stress is a major driving mechanism of COPD through the induction of chronic inflammation, induction of cellular senescence and impaired autophagy, reduced DNA repair, increased autoimmunity, increased mucus secretion, and impaired anti-inflammatory response to corticosteroids. Oxidative stress, therefore, drives the pathology of COPD and may increase disease progression, amplify exacerbations, and increase comorbidities through systemic oxidative stress. This suggests that antioxidants may be effective as disease-modifying treatments. Unfortunately, thiol-based antioxidants, such as N-acetylcysteine, have been poorly effective, as they are inactivated by oxidative stress in the lungs, so there is a search for more effective and safer antioxidants. New antioxidants in development include mitochondria-targeted antioxidants, NOX inhibitors, and activators of the transcription factor Nrf2, which regulates several antioxidant genes.

BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection of the respiratory system can progress to a multisystemic disease with aberrant inflammatory response. Cellular senescence promotes chronic inflammation, named senescence-associated secretory phenotype (SASP). We investigated whether coronavirus disease 2019 (COVID-19) is associated with cellular senescence and SASP.MethodsAutopsy lung tissue samples from 11 COVID-19 patients and 43 age-matched non-COVID-19 controls with similar comorbidities were analysed by immunohistochemistry for SARS-CoV-2, markers of senescence and key SASP cytokines. Virally induced senescence was functionally recapitulated in vitro, by infecting epithelial Vero-E6 cells and a three-dimensional alveosphere system of alveolar type 2 (AT2) cells with SARS-CoV-2 strains isolated from COVID-19 patients.ResultsSARS-CoV-2 …

Background: COPD is a disease of accelerated ageing, that may result from cellular senescence. MicroRNA-34a induces cellular senescence in small airway epithelial cells (SAEC) by inhibiting sirtuin-1 (SIRT1), an anti-ageing molecule. Extracellular vesicles (EVs) may contain microRNAs that can be modified in senescent cells. Senescent cells influence their environment and induce cellular senescence in neighbouring cells, thereby spreading senescence. The role of EVs in spreading senescence is currently unknown.Aim: To compare the effect of EVs produced by SAEC from COPD patients and non-smoker (NS) donors on cellular senescence of healthy SAECMethods: Primary SAEC were cultured in submerged culture. EVs were isolated by ultracentrifugation. Recipient cells were treated with EVs, and senescence markers were measured by RT-qPCR and Western blotResults: Healthy cells treated with …