ARL5b inhibits human rhinovirus 16 propagation and impairs macrophage-mediated bacterial clearance.

Human rhinovirus is the most frequently isolated virus during severe exacerbations of chronic respiratory diseases, like chronic obstructive pulmonary disease. In this disease, alveolar macrophages display significantly diminished phagocytic functions that could be associated with bacterial superinfections. However, how human rhinovirus affects the functions of macrophages is largely unknown. Macrophages treated with HRV16 demonstrate deficient bacteria-killing activity, impaired phagolysosome biogenesis, and altered intracellular compartments. Using RNA sequencing, we identify the small GTPase ARL5b to be upregulated by the virus in primary human macrophages. Importantly, depletion of ARL5b rescues bacterial clearance and localization of endosomal markers in macrophages upon HRV16 exposure. In permissive cells, depletion of ARL5b increases the secretion of HRV16 virions. Thus, we identify ARL5b as a novel regulator of intracellular trafficking dynamics and phagolysosomal biogenesis in macrophages and as a restriction factor of HRV16 in permissive cells.

Arpin is critical for phagocytosis in macrophages and is targeted by human rhinovirus.

Human rhinovirus is a causative agent of severe exacerbations of chronic obstructive pulmonary disease (COPD). COPD is characterised by an increased number of alveolar macrophages with diminished phagocytic functions, but how rhinovirus infection affects macrophage function is still unknown. Here, we describe that human rhinovirus 16 impairs bacterial uptake and receptor-mediated phagocytosis in macrophages. The stalled phagocytic cups contain accumulated F-actin. Interestingly, we find that human rhinovirus 16 downregulates the expression of Arpin, a negative regulator of the Arp2/3 complex. Importantly, re-expression of the protein rescues defective internalisation in human rhinovirus 16-treated cells, demonstrating that Arpin is a key factor targeted to impair phagocytosis. We further show that Arpin is required for efficient uptake of multiple targets, for F-actin cup formation and for successful phagosome completion in macrophages. Interestingly, Arpin is recruited to sites of membrane extension and phagosome closure. Thus, we identify Arpin as a central actin regulator during phagocytosis that it is targeted by human rhinovirus 16, allowing the virus to perturb bacterial internalisation and phagocytosis in macrophages.

The rheumatoid synovial environment alters fatty acid metabolism in human monocytes and enhances CCL20 secretion.

OBJECTIVES: Fatty acid oxidation (FAO) and glycolysis have been implicated in immune regulation and activation of macrophages. However, investigation of human monocyte intracellular metabolism in the context of the hypoxic and inflammatory rheumatoid arthritis (RA) synovium is lacking. We hypothesized that exposure of monocytes to the hypoxic and inflammatory RA environment would have a profound impact on their metabolic state, and potential to contribute to disease pathology. METHODS: Human monocytes were isolated from buffy coats and exposed to hypoxia. Metabolic profiling of monocytes was carried out by LC-MS metabolomics. Inflammatory mediator release after LPS or RA-synovial fluid (RA-SF) stimulation was analysed by ELISA. FAO was inhibited by etomoxir or enhanced with exogenous carnitine supplementation. Transcriptomics of RA blood monocytes and RA-SF macrophages was carried out by microarray. RESULTS: Hypoxia exacerbated monocyte-derived CCL20 and IL-1ß release in response to LPS, and increased glycolytic intermediates at the expense of carnitines. Modulation of carnitine identified a novel role for FAO in the production of CCL20 in response to LPS. Transcriptional analysis of RA blood monocytes and RA-SF macrophages revealed that fatty acid metabolism was altered and CCL20 increased when monocytes enter the synovial environment. In vitro analysis of monocytes showed that RA-SF increases carnitine abundance and CCL20 production in hypoxia, which was exacerbated by exogenous carnitine. CONCLUSION: This work has revealed a novel inflammatory mechanism in RA that links FAO to CCL20 production in human monocytes, which could subsequently contribute to RA disease pathogenesis by promoting the recruitment of Th17 cells and osteoclastogenesis.

MEK inhibition drives anti-viral defence in RV but not RSV challenged human airway epithelial cells through AKT/p70S6K/4E-BP1 signalling.

BACKGROUND: The airway epithelium is a major target tissue in respiratory infections, and its antiviral response is mainly orchestrated by the interferon regulatory factor-3 (IRF3), which subsequently induces type I (ß) and III (λ) interferon (IFN) signalling. Dual specificity mitogen-activated protein kinase kinase (MEK) pathway contributes to epithelial defence, but its role in the regulation of IFN response in human primary airway epithelial cells (AECs) is not fully understood. Here, we studied the impact of a small-molecule inhibitor (MEKi) on the IFN response following challenge with two major respiratory viruses rhinovirus (RV2) and respiratory syncytial virus (RSVA2) and a TLR3 agonist, poly(I:C). METHODS: The impact of MEKi on viral load and IFN response was evaluated in primary AECs with or without a neutralising antibody against IFN-ß. Quantification of viral load was determined by live virus assay and absolute quantification using qRT-PCR. Secretion of cytokines was determined by AlphaLISA/ELISA and expression of interferon-stimulated genes (ISGs) was examined by qRT-PCR and immunoblotting. A poly(I:C) model was also used to further understand the molecular mechanism by which MEK controls IFN response. AlphaLISA, siRNA-interference, immunoblotting, and confocal microscopy was used to investigate the effect of MEKi on IRF3 activation and signalling. The impact of MEKi on ERK and AKT signalling was evaluated by immunoblotting and AlphaLISA. RESULTS: Here, we report that pharmacological inhibition of MEK pathway augments IRF3-driven type I and III IFN response in primary human AECs. MEKi induced activation of PI3K-AKT pathway, which was associated with phosphorylation/inactivation of the translational repressor 4E-BP1 and activation of the protein synthesis regulator p70 S6 kinase, two critical translational effectors. Elevated IFN-ß response due to MEKi was also attributed to decreased STAT3 activation, which consequently dampened expression of the transcriptional repressor of IFNB1 gene, PRDI-BF1. Augmented IFN response translated into inhibition of rhinovirus 2 replication in primary AECs but not respiratory syncytial virus A2. CONCLUSIONS: Our findings unveil MEK as a key molecular mechanism by which rhinovirus dampens the epithelial cell's antiviral response. Our study provides a better understanding of the role of signalling pathways in shaping the antiviral response and suggests the use of MEK inhibitors in anti-viral therapy against RV.

Short Comprehensive Multimodal Behavioural Intervention for Tuberculosis-Can It Be Effectively Incorporated in Tribal Health?

BACKGROUND: Tuberculosis management in tribal areas is a major challenge to the National Tuberculosis Elimination Program in India. There is need for culturally appropriate interventions for bridging the gaps existing in the current system. There is paucity of research in this vulnerable group; hence, a study was undertaken to determine the effect of a Short Comprehensive Multimodal Behavioural Intervention in tribal colonies of Kerala. METHODS: The study used before-after design to assess the effectiveness of a Short Comprehensive Multimodal Behavioural Intervention for tuberculosis knowledge and voluntary reporting among residents of tribal colonies. The intervention included individual, small group, and large group education, with verbal, printed, and performance methods. Public-private partnership with community participation was emphasized to encourage the residents to approach public health system for managing tuberculosis. RESULTS: Ten tribal colonies from two districts were included with 104 participants. There was significant improvement in the proportion of participants with knowledge regarding different aspects of tuberculosis such as aetiology, symptoms, transmission, and treatment. The overall knowledge score had a significant improvement [median (range) 3.0 (0-9) to 7.0 (0-11), p < 0.001] when assessed one month consequent to the intense period of group education. CONCLUSION: Short-term health behavioural intervention package appropriate for the target group, implemented with public-private partnership and community participation of trained local volunteers, proved effective in improving the knowledge regarding tuberculosis and thereby health-seeking behaviour in detection. This can be tested for scaling up, and replication in other tribal health issues.

Effectiveness of physical stimulation for reducing injection pain in adults receiving intramuscular injections: a systematic review and meta-analysis.

OBJECTIVE: The objective of this review was to synthesize the best available research evidence regarding the effectiveness of physical stimulation for reducing injection pain in adults receiving intramuscular injections. INTRODUCTION: Pain associated with intramuscular injections continues to be a challenge for nurses. Various physical stimulation methods to alleviate pain and improve satisfaction for patients receiving intramuscular injections have been reported; however, the evidence surrounding the effectiveness of these methods remains inconclusive. INCLUSION CRITERIA: This systematic review considered randomized and quasi-experimental studies that used any physical stimulation strategies (eg, skin tapping, manual pressure, massage, pinch, traction) for adults aged 18 years and over receiving intramuscular injections. Studies that evaluated pain using validated instruments were considered for inclusion. METHODS: A three-step search strategy was conducted. MEDLINE, Embase, CINAHL, the Cochrane Library (Cochrane CENTRAL), Google Scholar, Dissertation Abstracts International, ProQuest Dissertations and Theses, and MedNar were searched from inception until 2020. We restricted the inclusion of studies to trials published in English. Two independent reviewers conducted the critical appraisal of eligible studies using the JBI checklists for randomized controlled and quasi-experimental trials. Data were extracted using the JBI data extraction tool, and meta-analysis and subgroup analysis were undertaken, where appropriate. RESULTS: Twenty-five studies were included with a total sample size of 1956 patients. Pooled results demonstrated that pain was significantly less with the use of the Helfer skin tap technique compared to no intervention (two studies; RR 0.73; 95% CI 0.66, 0.81; P <0.00001) or standard intervention (three studies; SMD -2.25; 95% CI -3.65, -0.85; P =0.002). Intervention with acupressure using standard treatment as control showed significant reduction in pain intensity (MD -4.78; 95% CI -5.32, -4.24; P <0.00001). Similarly, pain was significantly lower with manual pressure (two studies; SMD -0.42; 95% CI -0.69, 0.15; P =0.002) when compared to standard treatment. Pain scores were significantly lower in patients who received pinch technique, ShotBlocker, massage, or combination intervention (skin traction, pressure, and rapid muscle release) compared with no intervention, standard treatment, or placebo control. CONCLUSIONS: The evidence from this review demonstrates that physical stimulation - particularly the Helfer skin tap technique, acupressure, manual pressure, pinch technique, ShotBlocker, massage, and combination - can significantly lower intramuscular injection pain; however, this is based on low or very low certainty of evidence. SYSTEMATIC REVIEW REGISTRATION NUMBER: PROSPERO CRD42020168586.

Osimertinib + Savolitinib to Overcome Acquired MET-Mediated Resistance in Epidermal Growth Factor Receptor-Mutated, MET-Amplified Non-Small Cell Lung Cancer: TATTON.

MET-inhibitor and EGFR tyrosine kinase inhibitor (EGFR-TKI) combination therapy could overcome acquired MET-mediated osimertinib resistance. We present the final phase Ib TATTON (NCT02143466) analysis (Part B, n = 138/Part D, n = 42) assessing oral savolitinib 600 mg/300 mg once daily (q.d.) + osimertinib 80 mg q.d. in patients with MET-amplified, EGFR-mutated (EGFRm) advanced non-small cell lung cancer (NSCLC) and progression on prior EGFR-TKI. An acceptable safety profile was observed. In Parts B and D, respectively, objective response rates were 33% to 67% and 62%, and median progression-free survival (PFS) was 5.5 to 11.1 months and 9.0 months. Increased antitumor activity may occur with MET copy number ≥10. EGFRm circulating tumor DNA clearance on treatment predicted longer PFS in patients with detectable baseline ctDNA, while acquired resistance mechanisms to osimertinib + savolitinib were mediated by MET, EGFR, or KRAS alterations. SIGNIFICANCE: The savolitinib + osimertinib combination represents a promising therapy in patients with MET-amplified/overexpressed, EGFRm advanced NSCLC with disease progression on a prior EGFR-TKI. Acquired resistance mechanisms to this combination include those via MET, EGFR, and KRAS. On-treatment ctDNA dynamics can predict clinical outcomes and may provide an opportunity to inform earlier decision-making. This article is highlighted in the In This Issue feature, p. 1.

Exposure risk and COVID-19 infection among frontline health-care workers: A single tertiary care centre experience.

BACKGROUND: There is limited data on frontline health-care workers and risk of COVID-19 from the developing nations. It is imperative to identify those at higher risk to prevent further transmission. We assessed the relationship between exposure risk and COVID-19 among front-line health-care workers who were primary contacts of a COVID-19 patient. METHODS: A retrospective cohort study was conducted among front-line health-care workers in a tertiary care hospital who were exposed to a COVID-19 patient. Information on demographic factors, medical history, exposure related factors and subsequently COVID-19 lab reports were collected. An exposure risk assessment designed collating various exposure related factors categorized the participants into those with high and low risk. We used logistic regression to estimate the odds ratio of our primary outcome, a positive COVID-19 test when the independent variables were exposure risk, age, gender and occupation. RESULTS: Among1858 frontline workers who were primary contacts of a COVID-19 patient at the hospital, 106 (5.7%) incident reports of a positive COVID-19 test were recorded. None of the exposure related factors had any significant association with a positive COVID-19 test. However, high exposure risk category was significantly associated with COVID-19 positive test at the end of quarantine. CONCLUSION: COVID-19 was more frequent among front-line health-care workers who belonged to high exposure category. Education at different levels of service delivery at hospitals is required for best practice in order to prevent COVID-19 among health care providers. There is need to develop additional strategies to ensure that the information is translated in to practice.

A pilot study on glycemia and insulin resistance in patients with severe periodontitis.

CONTEXT: The potential impact of severe periodontitis on glycemia in systemically healthy individuals is not clearly established. It was hypothesized that among individuals who were previously undiagnosed for diabetes mellitus, patients with severe periodontitis have impaired glycemia and insulin resistance. AIMS: The aim of our study was to assess and compare glycemia in severe periodontitis patients and in individuals with clinically healthy periodontium. MATERIALS AND METHODS: A cross-sectional analytical design was employed. From among individuals who were undiagnosed for diabetes mellitus, 37 patients with severe periodontitis and 37 individuals with healthy periodontium in the age group of 25-55 years were recruited for the study. The fasting blood sugar (FBS), glycosylated hemoglobin (HbA1c), and insulin resistance by the Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) were assessed and compared between the two groups. RESULTS: The mean FBS, HOMA-IR, and HbA1c were significantly higher for patients with severe periodontitis than those individuals with healthy periodontium. After adjustments for age, gender, and body mass index, patients with severe periodontitis had a statistically significant association with impaired glucose metabolism (HbA1c ≥5.7) (adjusted odds ratio [OR] of 9.56; 95% confidence interval [CI]: 1.819-46.08; P < 0.01). Furthermore, patients with severe periodontitis had significantly greater odds to develop impaired fasting glucose (adjusted OR of 7.489, 95% CI: 1.408-39.839; P < 0.01). CONCLUSIONS: The mean FBS, HbA1c, and HOMA-IR were significantly higher in severe periodontitis patients than in the control group. A higher proportion of patients presented with prediabetes, incident diabetes, and insulin resistance in the severe periodontitis group.

Predicting Hospital Overall Quality Star Ratings in the USA.

The U.S. Centers for Medicare and Medicaid Services (CMS) assigns quality star ratings to hospitals upon assessing their performance across 57 measures. Ratings can be used by healthcare consumers for hospital selection and hospitals for quality improvement. We provide a simpler, more intuitive modeling approach, aligned with recent criticism by stakeholders. An ordered logistic regression approach is proposed to assess associations between performance measures and ratings across eligible (n = 4519) U.S. hospitals. Covariate selection reduces the double counting of information from highly correlated measures. Multiple imputation allows for inference of star ratings when information on all measures is not available. Twenty performance measures were found to contain all the relevant information to formulate star rating predictions upon accounting for performance measure correlation. Hospitals can focus their efforts on a subset of model-identified measures, while healthcare consumers can predict quality star ratings for hospitals ineligible under CMS criteria.

Effectiveness of physical stimulation on injection pain in adults receiving intramuscular injections: a systematic review protocol.

OBJECTIVE: The objective of this review is to evaluate the effectiveness of physical stimulation on injection pain in adults receiving intramuscular injections. INTRODUCTION: Intramuscular injections are the most commonly used modality for administration of pharmacological treatments. Despite this, pain from intramuscular injections is the most commonly reported side effect. Reducing patients' pain from intramuscular injections is important; however, the challenge is in selecting from the current methods available to alleviate pain, which are varied. The findings of this review may identify the most effective physical stimulation method to reduce the side effect of pain from an intramuscular injection. INCLUSION CRITERIA: This review will consider studies that include adults aged 18 years and over that use physical stimulation interventions during intramuscular injections. Any physical stimulation strategies used during intramuscular injections including devices, skin tapping, manual pressure, massage, pinch, and traction will be considered. Studies that evaluate pain using validated tools such as pain scales will be included. METHODS: The review will undertake to find both published and unpublished studies. The key information sources to be searched are MEDLINE, Embase, CINAHL, the Cochrane Library, Cochrane Central Register of Controlled Trials, Google Scholar, Dissertation Abstracts International, ProQuest Dissertations and Theses, and MedNar. Two independent reviewers will conduct a critical appraisal of eligible studies, assess the methodological quality, and extract the data. Studies will, where possible, be pooled in a statistical meta-analysis. SYSTEMATIC REVIEW REGISTRATION NUMBER: PROSPERO CRD42020168586.

Eosinophil-derived neurotoxin: A biologically and analytically attractive asthma biomarker.

There is a growing body of evidence for the utility of eosinophil-derived neurotoxin (EDN) as a biomarker in asthma, including association with eosinophilic airway inflammation, assessment of disease severity and potential for predicting pathogenic risks, including exacerbations. However, to interpret any biomarker data with confidence, it is first important to understand the preanalytical factors and biological variation that may affect its reliable measurement and results interpretation. In this study we defined the healthy serum EDN reference range for men and women as 1.98 to 26.10 ng/mL, with no significant gender differences. Smoking did not impact the mean EDN levels and no circadian rhythm was identified for EDN, unlike blood eosinophils (EOS) where levels peaked at 00:00h. EDN expression in different cell types was investigated and shown to occur primarily in eosinophils, indicating they are likely to be the main cellular repository for EDN. We also confirm that the quantification of serum EDN is not influenced by the type of storage tube used, and it is stable at ambient temperature or when refrigerated for at least 7 days and for up to one year when frozen at -20°C or -80°C. In summary, EDN is a stable biomarker that may prove useful in precision medicine approaches by enabling the identification of a subpopulation of asthma patients with activated eosinophils and a more severe form of the disease.

Full and partial agonists of muscarinic M3 receptors reveal single and oscillatory Ca2+ responses by beta 2-adrenoceptors.

Under physiological circumstances, cellular responses often reflect integration of signaling by two or more different receptors activated coincidentally or sequentially. In addition to heterologous desensitization, there are examples in which receptor activation either reveals or potentiates signaling by a different receptor type, although this is perhaps less well explored. Here, we characterize one such interaction between endogenous receptors in human embryonic kidney 293 cells in which Galpha(q/11)-coupled muscarinic M(3) receptors facilitate Ca(2+) signaling by Galpha(s)-coupled beta(2)-adrenoceptors. Measurement of changes in intracellular [Ca(2+)] demonstrated that noradrenaline released Ca(2+) from thapsigargin-sensitive intracellular stores only during activation of muscarinic receptors. Agonists with low efficacy for muscarinic receptor-mediated Ca(2+) responses facilitated cross-talk more effectively than full agonists. The cross-talk required Galpha(s) and was dependent upon intracellular Ca(2+) release channels, particularly inositol (1,4,5)-trisphosphate receptors. However, beta(2)-adrenoceptor-mediated Ca(2+) release was independent of measurable increases in phospholipase C activity and resistant to inhibitors of protein kinases A and C. Interestingly, single-cell imaging demonstrated that particularly lower concentrations of muscarinic receptor agonists facilitated marked oscillatory Ca(2+) signaling to noradrenaline. Thus, activation of muscarinic M(3) receptors profoundly influences the magnitude and oscillatory behavior of intracellular Ca(2+) signaling by beta(2)-adrenoceptors. Although these receptor subtypes are often coexpressed and mediate contrasting acute physiological effects, altered oscillatory Ca(2+) signaling suggests that cross-talk could influence longer term events through, for example, regulating gene transcription.

Dual Role For A MEK Inhibitor As A Modulator Of Inflammation And Host Defense Mechanisms With Potential Therapeutic Application In COPD.

Background: Unlike p38 mitogen-activated protein Kinases (MAPK) that has been extensively studied in the context of lung-associated pathologies in COPD, the role of the dual-specificity mitogen-activated protein kinase kinase (MEK1/2) or its downstream signaling molecule extracellular signal-regulated kinases 1/2 (ERK1/2) in COPD is poorly understood. Objectives: The aim of this study was to address whether MEK1/2 pathway activation is linked to COPD and that targeting this pathway can improve lung inflammation through decreased immune-mediated inflammatory responses without compromising bacterial clearance. Methods: Association of MEK1/2 pathway activation to COPD was investigated by immunohistochemistry using lung tissue biopsies from COPD and healthy individuals and through analysis of sputum gene expression data from COPD patients. The anti-inflammatory effect of MEK1/2 inhibition was assessed on cytokine release from lipopolysaccharide-stimulated alveolar macrophages. The effect of MEK1/2 inhibition on bacterial clearance was assessed using Staphylococcus aureus killing assays with RAW 264.7 macrophage cell line and human neutrophils. Results: We report here MEK1/2 pathway activation demonstrated by increased pERK1/2 staining in bronchial epithelium and by the presence of MEK gene activation signature in sputum samples from COPD patients. Inhibition of MEK1/2 resulted in a superior anti-inflammatory effect in human alveolar macrophages in comparison to a p38 inhibitor. Furthermore, MEK1/2 inhibition led to an increase in bacterial killing in human neutrophils and RAW 264.7 cells that was not observed with the p38 inhibitor. Conclusion: Our data demonstrate the activation of MEK1/2 pathway in COPD and highlight a dual function of MEK1/2 inhibition in improving host defense responses whilst also controlling inflammation.

Synthesis of 1,2,3-triazole-linked pyrrolobenzodiazepine conjugates employing 'click' chemistry: DNA-binding affinity and anticancer activity.

1,2,3-Triazole based molecules are useful pharmacophores for several DNA-alkylating and cross-linking agents. A series of A/C8, C/C2 and A/C8-C/C2-linked 1,2,3-triazole-PBD conjugates have been synthesized by employing 'click' chemistry. These molecules have exhibited promising DNA-binding affinity and anticancer activity in selected human cancer cell lines.

Phosphonate-linked pyrrolo[2,1-c][1,4]benzodiazepine conjugates: synthesis, DNA-binding affinity and cytotoxicity.

Pyrrolobenzodiazepine-diethylphosphonate conjugates have been designed and synthesized that link through two different types of spacers that are simple alkane chain and also a piperazine moiety side-armed with the alkane chains. These pyrrolobenzodiazepine conjugates have exhibited remarkable DNA-binding affinity and improved solubility in water, a representative compound 7d showing promising in vitro cytotoxicity.

Synthesis, DNA-binding ability and evaluation of antitumour activity of triazolo[1,2,4]benzothiadiazine linked pyrrolo[2,1-c][1,4]benzodiazepine conjugates.

A series of triazolobenzothiadiazine-pyrrolobenzodiazepine conjugates linked through different alkane spacers have been prepared. These compounds have exhibited significant cytotoxicity against most of the cell lines examined. Compound 5a displays GI(50) values from 1.83 to 2.38 microM against seven human tumour cell lines, and is identified as a promising lead compound from this series. Their DNA thermal denaturation studies have also been carried out, and one of the compounds 5c elevates the DNA helix melting temperature of the CT-DNA by 2.6 degrees C after incubation for 36 h.

HRV16 Impairs Macrophages Cytokine Response to a Secondary Bacterial Trigger.

Human rhinovirus is frequently seen as an upper respiratory tract infection but growing evidence proves the virus can cause lower respiratory tract infections in patients with chronic inflammatory lung diseases including chronic obstructive pulmonary disease (COPD). In addition to airway epithelial cells, macrophages are crucial for regulating inflammatory responses to viral infections. However, the response of macrophages to HRV has not been analyzed in detail. We used in vitro monocyte-derived human macrophages to study the cytokine secretion of macrophages in response to the virus. Our results showed that macrophages were competent at responding to HRV, as a robust cytokine response was detected. However, after subsequent exposure to non-typeable Haemophilus influenzae (NTHi) or to LPS, HRV-treated macrophages secreted reduced levels of pro-inflammatory or regulatory cytokines. This "paralyzed" phenotype was not mimicked if the macrophages were pre-treated with LPS or CpG instead of the virus. These results begin to deepen our understanding into why patients with COPD show HRV-induced exacerbations and why they mount a defective response toward NTHi.

The development of AZD7624 for prevention of exacerbations in COPD: a randomized controlled trial.

Background: p38 mitogen-activated protein kinase (MAPK) plays a central role in the regulation and activation of pro-inflammatory mediators. COPD patients have increased levels of activated p38 MAPK, which correlate with increased lung function impairment, alveolar wall inflammation, and COPD exacerbations. Objectives: These studies aimed to assess the effect of p38 inhibition with AZD7624 in healthy volunteers and patients with COPD. The principal hypothesis was that decreasing lung inflammation via inhibition of p38α would reduce exacerbations and improve quality of life for COPD patients at high risk for acute exacerbations. Methods: The p38 isoform most relevant to lung inflammation was assessed using an in situ proximity ligation assay in severe COPD patients and donor controls. Volunteers aged 18-55 years were randomized into the lipopolysaccharide (LPS) challenge study, which investigated the effect of a single dose of AZD7624 vs placebo on inflammatory biomarkers. The Proof of Principle study randomized patients aged 40-85 years with a diagnosis of COPD for >1 year to AZD7624 or placebo to assess the effect of p38 inhibition in decreasing the rate of exacerbations. Results: The p38 isoform most relevant to lung inflammation was p38α, and AZD7624 specifically inhibited p38α and p38ß isoforms in human alveolar macrophages. Thirty volunteers were randomized in the LPS challenge study. AZD7624 reduced the increase from baseline in sputum neutrophils and TNF-α by 56.6% and 85.4%, respectively (p<0.001). In the 213 patients randomized into the Proof of Principle study, there was no statistically significant difference between AZD7624 and placebo when comparing the number of days to the first moderate or severe exacerbation or early dropout. Conclusion: Although p38α is upregulated in the lungs of COPD patients, AZD7624, an isoform-specific inhaled p38 MAPK inhibitor, failed to show any benefit in patients with COPD.