NIHRDataInsights/HRCSData huggingface dataset:
Title: Pro-inflammatory lung dendritic cells in stratified severe RSV bronchiolitis
Funder: Medical Research Council

Abstract: RSV is the main cause of viral bronchiolitis in infants worldwide. With significant morbidity and mortality RSV bronchiolitis has major health and economic implications. It results in hospitalisation of 2% of all infants who may develop severe disease with respiratory failure requiring mechanical ventilation. While for most this is only required for a few days, in a third of very severe cases resolution of disease is delayed necessitating prolonged mechanical ventilation. Respiratory disease in RSV bronchiolitis is due to airflow obstruction resulting from excessive inflammation of the small terminal airways. The immune mechanisms by which this inflammation is maintained, delaying resolution of disease, are not known. Mouse models of RSV infection suggest important roles of the expanded population of pro-inflammatory conventional dendritic cells (cDCs) and associated T cell responses. We have recently detected sizable populations of activated cDCs and T cells in bronchoalveolar lavages (BAL) from infants with severe RSV disease. Here, we will test the hypothesis that activated lung cDCs and associated pro-inflammatory T cell responses are essential for the maintenance of airway inflammation in prolonged severe RSV bronchiolitis. Using a clinical study comparing infants with faster or delayed resolution of RSV disease, we will determine cDC numbers in BAL, their activation status and their ability to induce pro-inflammatory T cell responses, and we will assess cytokine concentrations in BAL fluid and their ability to differentiate and activate cDCs. Using the mouse model of RSV infection, we will define the role of cDCs for lung inflammation and disease by depletion and reconstitution. These translational studies will define the role of cDCs and associated T cell responses for the maintenance of inflammation in delayed resolution of severe RSV bronchiolitis and may identify cDCs as a new treatment target and biomarker in this condition.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: RESPIRATORY
Title: Myofibroblast differentiation in idiopathic pulmonary fibrosis: epigenetic mechanisms and therapeutic targeting by cAMP
Funder: Medical Research Council

Abstract: Idiopathic pulmonary fibrosis (IPF) is a deadly respiratory disease of unknown aetiology with no effective therapy. IPF is characterised by fibroblast accumulation and activation and excessive collagen deposition, leading to distortion of the alveolar architecture, progressive loss of lung function, and ultimately death. Myofibroblasts are the primary effector cells in IPF and can be differentiated from lung fibroblasts and epithelial cells. PGE2, a major prostanoid generated by cyclooxygenase-2 (COX-2) from lung fibroblasts and epithelial cells, inhibits myofibroblast differentiation through cAMP. However, PGE2 is reduced due to COX-2 repression in this process, leading to unopposed myofibroblast differentiation. Increasing evidence suggests that epigenetic regulation plays a key role in myofibroblast differentiation and that cAMP elevating agents can compensate for exogenous PGE2 loss to prevent and even reverse myofibroblast differentiation. We hypothesize that lung fibroblasts and epithelial cells are epigenetically reprogramed during their differentiation to myofibroblasts in IPF, resulting in gene specific epigenetic modifications relative to the expression and repression key genes and that cAMP elevating agents can prevent, even reverse, the differentiation at least partly through epigenetic regulation. We aim to elucidate the epigenetic mechanisms and cAMP-mediated inhibition of myofibroblast differentiation by studying TGF-beta1-induced fibroblast-to-myofibrobalst differentiation and epithelial-mesenchymal transition in vitro, validating key epigenetic findings relative to gene expression in IPF lung tissue and in a mouse model of pulmonary fibrosis, and evaluating the anti-fibrotic effect of cAMP elevating agents and epigenetic inhibitors in the mouse model. The outcome of this project will improve our understanding of IPF pathogenesis and provide preclinical evidence for these agents as potential novel therapies to benefit IPF patients.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: RESPIRATORY
Title: Effect of exposure of children to particulate matter air pollution on airway macrophages and dendritic cells.
Funder: Medical Research Council

Abstract: We will determine the association between chronic exposure of children to PM air pollution and; i) the proportion of small airway macrophages (AM), ii) the number, proportion and activation status of dendritic cells with a mature phenotype, and iii) the capacity of dendritic cells to stimulate naïve CD4+ T cells in vivo. Using AM carbon as a biomarker of individual exposure to PM, we will test the hypothesis that PM, by stimulating mediator release by lung cells, enhances the recruitment of the myeloid precursors of AM and dendritic cells, and directly stimulating expression of co-stimulatory receptors on dendritic cells, results in an airway milieu with an increased proportion of smaller "pro-inflammatory' AM, and a dendritic cell phenotype with increased capacity to stimulate CD4+ T cell proliferation and generate Th2 responses. To assess this, we will recruit 400 children via the Centre of the Cell resource - a multimedia teaching experience embedded in our Institute. Lung function, atopic status, exhaled breath condensate pro-inflammatory mediators, and glutathione gene polymorphism status (GSTP, GSTM) will be done in all children. The local sources of PM exposure will be assessed in all children using personal black carbon monitoring on a typical school day. Lower airway cells will be sampled using induced sputum. AM carbon will be determined using our established image analysis method. Markers for recruitment of myeloid precursors and co-stimulatory molecule expression will be assessed by flow cytometry. In functional studies, we will assess the capacity of induced sputum cells to stimulate proliferation of naïve CD4+ T cells proliferation. The study is powered to detect a 0.6 SD in the primary outcome variables between children in the upper and lower tertiles of airway macrophage carbon. All the sampling procedures to be used have been ethically approved in children, and our flow cytometry analysis standard operating procedure is based on robust pilot data.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;2.2 FACTORS RELATING TO PHYSICAL ENVIRONMENT
Health categories: RESPIRATORY
Title: MICA: Evaluation of anti diabetic drugs in the treatment of respiratory disease
Funder: Medical Research Council

Abstract: The respiratory tract is lined with a thin layer of fluid (airway surface liquid, ASL), which is important for lung defence against infection. Glucose concentrations in ASL are normally 12x lower than that of blood. However, they are raised in people during airway inflammation, hyperglycaemia and, more potently, when both are present, making the airway more susceptible to infection from pathogens such as methicillin-resistant S.aureus and P.aeruginosa. We have shown in airway epithelial cells in vitro that, under normal conditions, glucose diffuses from blood into ASL but accumulation is limited by the permeability of the epithelium, blood glucose concentration, glucose uptake into the epithelial cell via GLUT transporters and its metabolism. Inflammation increased transepithelial permeability to glucose and elevated glucose in ASL. In addition, increasing basolateral glucose concentration increased glucose in ASL and promoted the growth of S.aureus on the apical surface of lung epithelial cells. In vitro, pre-treatment of airway epithelia with metformin (a drug used in the treatment of type II diabetes) reduced epithelial permeability to glucose and prevented the apical growth of S. aureus. Metformin had no direct effect on bacterial growth or on basolateral glucose concentration. This project will investigate how lung glucose homeostasis is modified by inflammation and hyperglycaemia in vitro and in vivo. It will identify whether metformin inhibits glucose accumulation in ASL and limits the growth of respiratory pathogens in vivo. It will also investigate the therapeutic effect of other drugs which are reported to reduce epithelial permeability or reduce blood glucose concentrations. e.g. azithromycin, glucokinase activators and SGLT2 inhibitors. The outcomes of this project will determine whether these drugs have potential therapeutic benefit in the prevention and treatment of respiratory infection and chronic pulmonary disease.


Research activity codes: 2.2 FACTORS RELATING TO PHYSICAL ENVIRONMENT;5.1 PHARMACEUTICALS
Health categories: RESPIRATORY
Title: A new paradigm for testing pathway tractability in lung disease
Funder: Medical Research Council

Abstract: The archetypal pathway of drug validation utilises cell-based assays, an animal model of disease followed by phased studies in man. Pre-clinical models are of little value in predicting clinical efficacy, and many drugs are entered into clinical trial without 'proof' that they are biologically effective against their intended target in the disease itself. We describe a novel tractable strategy for optimising the pathway of drug development using specific optical Smartprobes and direct visualisation of pathway activity/inhibition in situ in real time using probe-based confocal laser endomicroscopy (pCLE). This pathway incorporates clear go/no go steps to reduce attrition rate, enrich the pool of drugs that progress to efficacy studies, and offers a mechanism-based method for stratifying patients for drug trials in diseases of the lung and indeed any organ accessible by fibreoscopy. Using idiopathic pulmonary fibrosis (IPF), a devastating disease, as the model, and the MMP9/12 inhibitor AZD1236 as the drug, we aim to quantify the real-time intrapulmonary activity of MMP9 and drug-mediated MMP9 inhibition in patients with IPF. Methodology 1. In vitro evaluation of drug-mediated MMP9 activity inhibition utilising a MMP9-substrate Smartprobe and fluoro-labelled AZD1236 in cells and fresh tissue from patients with IPF. These studies aim to show colocalisation of the drug and Smartprobe and real-time inhibition of Smartprobe-activity. 2. Microdosing studies of AZD1236 and MMP9 Smartprobe in man. This includes a first-in-man study of dual microdosed drug and Smartprobe followed by an experimental study of intrapulmonary drug and Smartprobe in patients with IPF. The study is designed to prove, in situ real-time, drug-mediated MMP9 inhibition in fibrotic lung. 3. A phase IIa double blinded RCT study of AZD1236 given by the intended route (oral) utilising MMP9 Smartprobe and pCLE in IPF. Primary end-point is change in intrapulmonary MMP9 Smartprobe activity at 6 weeks.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: RESPIRATORY
Title: Resolving mediators as novel therapies for idiopathic pulmonary fibrosis
Funder: Medical Research Council

Abstract: General experimental protocols: Primary human lung myofibroblasts (PHLMF) are grown from the lung parenchyma of patients with IPF and healthy non-fibrotic control tissue removed at lung resection. Human fibrocytes are grown from peripheral blood progenitors. Human lung explant tissue is obtained from healthy areas of lung removed during surgery for carcinoma. AIM 1: To examine the effect of resolving mediators on PHLMF and fibroblast progenitors (peripheral blood fibrocytes) collected from patients with IPF compared to non fibrotic controls. We will investigate: a) The effects of LXA4 and RvD2 on PHLMF biology (FBS and basic FGF-dependent wound healing, proliferation, TGFbeta-dependent collagen secretion, constitutive alphaSMA expression, contraction). b) The effects of LXA4 and RvD2 on primary human fibrocyte biology using established techniques as described previously (wound healing, collagen secretion, differentiation, migration). c) Whether resolving mediators inhibit epithelial-mesenchymal transition (EMT) using immunofluorescence staining on epithelial cell lines and primary epithelial cells. d) Whether resolving mediators inhibit TGFbeta1-dependent cell signaling in both PHLMF and epithelial cells using various complementary techniques including Western blot and ELISA assays, immunohistochemistry, flow cytometry, and a luciferase reporter construct. e) Assessment of resolving mediator receptor expression on PHLMF and fibrocytes using real-time RT-PCR, flow cytometry or Western blot. AIM 2: To examine the effect of resolving mediators on TGFbeta-dependent responses in ex vivo human lung parenchyma (collagen secretion, alphaSMA expression) using immunohistochemistry and the measurement of soluble mediators in the culture supernatant. AIM 3: To measure the concentrations of resolving mediators in the BAL of IPF patients compared to healthy age and smoking matched controls using commercial ELISAs and liquid chromatography-mass spectrometry


Research activity codes: 5.1 PHARMACEUTICALS;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: RESPIRATORY
Title: MICA - Investigation into the mechanisms that drive cough in health and disease
Funder: Medical Research Council

Abstract: The goal of this collaboration would be to provide insight into the mechanisms that drive cough in health and disease and to understand the similarities and differences in exaggerated cough that is present in specific patient groups. This would provide novel target validation data and inform a translational and predictive screening pathway from high throughput assays to clinical trial design. (1) Investigate which endogenous mediators (produced in inflammatory diseases) and environmental irritants are able to elicit cough, which fibre type they activate and evaluate their mechanism of action. (2) Evaluate the utility of selective ligands in cell lines overexpressing the corresponding target protein. Establish which TRP channels are involved in the activation of airway sensory nerves and evoke a cough reflex under 'normal' conditions using selective ligands and vagal tissue from gene deleted animals (eg. TRPV1, TRPV4, TRPA1). If Ad and C-fibre afferents are involved preferentially evaluate the role of NaV channels (in particular NaV1.7). We will perform pharmacokinetic/pharmacodynamic (PK/PD) modelling integrating compound exposure information with in vitro and in vivo data which will aid in the interpretation of the data sets. (3) Investigate changes in the function and phenotype of airway specific afferents and cough in disease models (eg. allergen, sensitised and challenged guinea-pigs; CS exposed guinea-pigs) as previously described by our lab and others using selective ligands. Develop a model of post viral cough (mirroring the most appropriate clinical phenotype eg. H1N1, rhinovirus). (4) Perform clinical studies using ion channel specific tussive stimuli (eg. TRPV1, capsaicin; TRPA1, cinnamaldehyde; TRPV4, investigate appropriate stimuli, our pre-clinical studies suggest hypertonic saline may partially activate through TRPV4) across the relevant patient groups (normal, non-smokers, smokers, asthmatics, COPD, idiopathic coughers).


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: RESPIRATORY
Title: The role of hypoxia in determining the cellular outcomes of pulmonary infection and its importance in the pathogenesis of COPD
Funder: Medical Research Council

Abstract: COPD represents a significant burden of disease world wide, yet the cellular pathways determining disease progression remain poorly understood. It is recognised however, that patients who have frequent infective exacerbations of respiratory symptoms have a worse prognosis and that abnormal gas exchange is often observed either acutely during an exacerbation or chronically as the disease progresses. The role of systemic hypoxia in determining disease progression, susceptibility to pulmonary bacterial infections and persistent inflammatory responses has yet to be addressed. The key goals of my research proposal therefore are to: 1. Characterise the effects of systemic hypoxia on outcomes of acute pulmonary infections using murine models of bacterial pneumonia. 2. Investigate the role of HIF oxygen sensing pathway members in determining bacterial clearance and myeloid cell mediated inflammation in acute pulmonary infections. 3. Define whether systemic administration of hydroxylase inhibitors can modify susceptibility to and outcomes of acute bacterial pneumonia in mice in the context of hypoxia. 4. Determine whether ventilatory failure and infective exacerbations modify neutrophil expression of HIFs and dioxygenase family members in patients with COPD. Through a better understanding of these oxygen-regulated signalling pathways, I hope to identify specific targets with potential to limit inappropriate inflammation whilst preserving anti-microbial responses.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: RESPIRATORY
Title: Optimisation of scaffold matrix for epithelial regeneration on tissue engineered airway implants.
Funder: Medical Research Council

Abstract: Aim - Optimisation of scaffold matrix for epithelial regeneration on tissue engineered airway implants Objectives 1. Determine the key extracellular matrix protein(s) important in the regeneration of respiratory epithelium from a selection of candidate proteins already identified 2. Examine if key ECM protein(s) can be used to enhance respiratory epithelial regeneration on synthetic and collagen scaffolds. 3. To investigate the role of the beta1 and alpha6 integrin subunits in cell adherence and survival of seeded airway epithelium and the downstream signalling pathways responsible Methodology i) To define the role of key ECM proteins in cell adhesion, survival, proliferation and differentiation using 2D and 3D organoid culture systems ii) To delineate the effects of protein coating of target ECM proteins on epithelialisation of POSS-PCU using air liquid interface culture systems iii) Examine the use of compressed collagen sheets in epithelial cell preparation using air liquid interface culture systems iv) Examine integrin ECM protein interactions using blocking antibodies and retroviral transduction to overexpress beta one and alpha six integrin receptors. Scientific and medical opportunities a) Improved understanding of respiratory epithelial homeostasis and repair b) Improved respiratory epithelial regeneration in-vitro on POSS-PCU to improve the safety and effectiveness of tissue engineered airway implants c) Production of respiratory epithelial collagen sheets that would open up new therapeutic options for those with chronic upper airway disease.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;5.2 CELLULAR AND GENE THERAPIES
Health categories: RESPIRATORY
Title: Lung health and exposure to household air pollution in rural Malawi
Funder: Medical Research Council

Abstract: Design: 1) An exposure-incidence study will be conducted within CAPS to determine the association between exposure to household air pollution (carbon monoxide [CO]) and pneumonia in children under the age of 5 in rural Malawi. 2) A cross-sectional study will be conducted to determine the prevalence and determinants of obstructive lung disease in adults in rural Malawi. 3) A prospective cohort study will be conducted within CAPS to determine the extent to which exposure to CO and particulate matter (PM2.5) explains the rate of decline in lung function in adults in rural Malawi. Study population: Exposure-incidence study - Children up to 5 years old included in CAPS Cross-sectional and cohort studies - Non-institutionalised adults aged 18 and above Exposures: CO (Lascar CO monitors) and particulate matter (UCB particle monitors) exposures from biomass smoke Outcomes: Exposure-incidence study - Pneumonia in children under 5 years of aged diagnosed by blinded physicians or medical officers using the WHO Integrated Management of Childhood Illness (IMCI) pneumonia assessment protocol Cross-sectional and cohort studies - Post bronchodilator spirometry in adults Sample size and potential power: Exposure-incidence study - With 2000 children this study will have 90% power to detect a minimum mean difference between the controls and cases of 6.53 (40%) ppm CO. Cross-sectional and cohort study - With 2000 adults these studies will provide an estimate of prevalence of acceptable precision (e.g. with an estimated prevalence of 15% the estimated 95% CI for each gender would be 15% +/- 4%) and have 90% power to detect a correlation between particulate matter exposure and change in FEV1 level of 0.102 (or greater) in both age group combined and 0.144 (or greater) in each age group separately. Consumer involvement: Villagers, local stove producers and community leaders have been involved with the development of this work.


Research activity codes: 2.2 FACTORS RELATING TO PHYSICAL ENVIRONMENT;2.4 SURVEILLANCE AND DISTRIBUTION
Health categories: RESPIRATORY
Title: Analysis of the pulmonary GR interactome, and functional validation.
Funder: Medical Research Council

Abstract: Lung disease, frequently with a major inflammatory component, is a major human health burden, in terms of cost, and morbidity/mortality. Glucocorticoids are widely used to treat lung inflammation, but the treatment response is variable. In addition some inflammatory diseases do not respond to glucocorticoids. We have developed novel approaches to comprehensively measure the GR post-translational modification, and protein interactome in lung tissue. This permits, for the first time, sampling of the consequences of inflammation, and drug treatment within the target tissue under physiological conditions. As a consequence of our pilot work we have already discovered an unexpected role for the protein caveolin as a GR regulator, and seek to explore the mechanisms responsible at an early stage. We have preliminary data associating changes in pulmonary inflammatory reactivity with circulating endogenous glucocorticoids, and therefore also seek to explore how these two systems interact, using in-vivo and in-vitro approaches. We move to cell-type specific targeting of GR expression to anchor our observations to the bronchial epithelial cells, permitting further proteomics analysis in defined cell populations from whole animal challenges. We will explore the mechanisms of GR regulation, and the consequences for GR function using cell models, and molecular analysis at two levels, the GR cistrome and transcriptome. We have unrivalled expertise in computational biology required to manage these multidimensional datasets at applicant, and collaborator level, and outcomes will include both systems level analysis, and the generation of testable hypotheses. These latter will be pursued using cell culture models, before return to experimental pulmonary inflammation. We will translate our findings to human lung disease by associating changes in emergent target gene expression, and protein in our comprehensive lung tissue archive.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: RESPIRATORY