82 results found
Cheong S-S, Luis T, Stewart M, et al., 2023, TReATS: a novel method for TAT-Cre recombinase mediated floxed Allele modification in ex vivo tissue slices, Disease Models and Mechanisms, Vol: 16, ISSN: 1754-8403
Precision-Cut Lung Slices (PCLS) are used for a variety of applications. However, methods to manipulate genes in PCLS are currently limited. We developed a novel method, TAT-Cre Recombinase-mediated floxed Allele modification in Tissue Slices (TReATS), to induce highly effective and temporally controlled gene deletion or activation in ex vivo PCLS. Treatment of PCLS from Rosa26-flox-stop-flox-EYFP mice with cell-permeant TAT-Cre recombinase induced ubiquitous EYFP protein expression, indicating successful Cre-mediated excision of the upstream loxP-flanked stop sequence. Quantitative real-time PCR confirmed induction of EYFP. We successfully replicated the TReATS method in PCLS from Vangl2flox/flox mice, leading to the deletion of loxP-flanked exon 4 of the Vangl2 gene. Cre-treated Vangl2flox/flox PCLS exhibited cytoskeletal abnormalities, a known phenotype caused by VANGL2 dysfunction. We report a novel method that by-passes conventional Cre-Lox breeding, allowing rapid and highly effective gene manipulation in ex vivo tissue models.
Buttery SC, Banya W, Bilancia R, et al., 2023, Lung volume reduction surgery versus endobronchial valves: a randomised controlled trial, European Respiratory Journal, Vol: 61, Pages: 1-14, ISSN: 0903-1936
BACKGROUND: Lung volume reduction surgery (LVRS) and bronchoscopic lung volume reduction (BLVR) with endobronchial valves can improve outcomes in appropriately selected patients with emphysema. However, no direct comparison data exist to inform clinical decision making in people who appear suitable for both procedures. Our aim was to investigate whether LVRS produces superior health outcomes when compared with BLVR at 12 months. METHODS: This multicentre, single-blind, parallel-group trial randomised patients from five UK hospitals, who were suitable for a targeted lung volume reduction procedure, to either LVRS or BLVR and compared outcomes at 1 year using the i-BODE score. This composite disease severity measure includes body mass index, airflow obstruction, dyspnoea and exercise capacity (incremental shuttle walk test). The researchers responsible for collecting outcomes were masked to treatment allocation. All outcomes were assessed in the intention-to-treat population. RESULTS: 88 participants (48% female, mean±sd age 64.6±7.7 years, forced expiratory volume in 1 s percent predicted 31.0±7.9%) were recruited at five specialist centres across the UK and randomised to either LVRS (n=41) or BLVR (n=47). At 12 months follow-up, the complete i-BODE was available in 49 participants (21 LVRS/28 BLVR). Neither improvement in the i-BODE score (LVRS -1.10±1.44 versus BLVR -0.82±1.61; p=0.54) nor in its individual components differed between groups. Both treatments produced similar improvements in gas trapping (residual volume percent predicted: LVRS -36.1% (95% CI -54.6- -10%) versus BLVR -30.1% (95% CI -53.7- -9%); p=0.81). There was one death in each treatment arm. CONCLUSION: Our findings do not support the hypothesis that LVRS is a substantially superior treatment to BLVR in individuals who are suitable for both treatments.
Ferreira PM, Bayer S, Zhu D, et al., 2022, Extracellular Vesicles in Lung Diseases, EXTRACELLULAR VESICLES, Editors: Chrzanowski, Lim, Kim, Publisher: ROYAL SOC CHEMISTRY, Pages: 216-245, ISBN: 978-1-78801-894-4
Roisin M, Kim SY, Van der Plaat D, et al., 2021, LSC-2021-Investigating the role of vitamin A intake and retinoic acid signalling in lung homeostasis and repair-A multidisciplinary approach, Publisher: EUROPEAN RESPIRATORY SOC JOURNALS LTD, ISSN: 0903-1936
Hind M, Wong T, 2021, Atrial Fibrillation, Obstructive Sleep Apnea, and Continuous Positive Airway Pressure: No Easy Fix, AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, Vol: 204, Pages: 503-505, ISSN: 1073-449X
Bush A, Pabary R, Allinson J, et al., 2021, They SHALL grow old: a UK rare disease clinical network for adult congenital thoracic malformations, ARCHIVES OF DISEASE IN CHILDHOOD, Vol: 106, Pages: 625-626, ISSN: 0003-9888
Kim SY, Mongey S, Wang P, et al., 2021, The Acid Injury and Repair (AIR) model: A new ex vivo tool to understand lung repair, Biomaterials, Vol: 267, ISSN: 0142-9612
Research into mechanisms underlying lung injury and subsequent repair responses is currently of paramount importance. There is a paucity of models that bridge the gap between in vitro and in vivo research. Such intermediate models are critical for researchers to decipher the mechanisms that drive repair and to test potential new treatments for lung repair and regeneration. Here we report the establishment of a new tool, the Acid Injury and Repair (AIR) model, that will facilitate studies of lung tissue repair. In this model, injury is applied to a restricted area of a precision-cut lung slice using hydrochloric acid, a clinically relevant driver. The surrounding area remains uninjured, thus mimicking the heterogeneous pattern of injury frequently observed in lung diseases. We show that in response to injury, the percentage of progenitor cells (pro surfactant protein C, proSP-C and TM4SF1 positive) significantly increases in the injured region. Whereas in the uninjured area, the percentage of proSP-C/TM4SF1 cells remains unchanged but proliferating cells (Ki67 positive) increase. These effects are modified in the presence of inhibitors of proliferation (Cytochalasin D) and Wnt secretion (C59) demonstrating that the AIR model is an important new tool for research into lung disease pathogenesis and potential regenerative medicine strategies.
Kim S, Mongey R, Griffiths M, et al., 2020, An ex vivo acid injury and repair (AIR) model using precision-cut lung slices to understand lung injury and repair, Current protocols in mouse biology, Vol: 10, Pages: e85-e85, ISSN: 2161-2617
Recent advances in cell culture models like air‒liquid interface culture and ex vivo models such as organoids have advanced studies of lung biology; however, gaps exist between these models and tools that represent the complexity of the three‐dimensional environment of the lung. Precision‐cut lung slices (PCLS) mimic the in vivo environment and bridge the gap between in vitro and in vivo models. We have established the acid injury and repair (AIR) model where a spatially restricted area of tissue is injured using drops of HCl combined with Pluronic gel. Injury and repair are assessed by immunofluorescence using robust markers, including Ki67 for cell proliferation and prosurfactant protein C for alveolar type 2/progenitor cells. Importantly, the AIR model enables the study of injury and repair in mouse lung tissue without the need for an initial in vivo injury, and the results are highly reproducible. Here, we present detailed protocols for the generation of PCLS and the AIR model. We also describe methods to analyze and quantify injury in AIR‐PCLS by immunostaining with established early repair markers and fluorescence imaging. This novel ex vivo model is a versatile tool for studying lung cell biology in acute lung injury and for semi‐high‐throughput screening of potential therapeutics. © 2020 Wiley Periodicals LLC.
Cheong SS, Akram K, Metellan C, et al., 2020, The planar polarity component Vangl2 is a key regulator of mechanosignaling, Frontiers in Cell and Developmental Biology, Vol: 8, ISSN: 2296-634X
VANGL2 is a component of the planar cell polarity (PCP) pathway, which regulates tissue polarity and patterning. The Vangl2Lp mutation causes lung branching defects due to dysfunctional actomyosin-driven morphogenesis. Since the actomyosin network regulates cell mechanics, we speculated that mechanosignaling could be impaired when VANGL2 is disrupted. Here, we used live-imaging of precision-cut lung slices (PCLS) from Vangl2Lp/+ mice to determine that alveologenesis is attenuated as a result of impaired epithelial cell migration. Vangl2Lp/+ tracheal epithelial cells (TECs) and alveolar epithelial cells (AECs) exhibited highly disrupted actomyosin networks and focal adhesions (FAs). Functional assessment of cellular forces confirmed impaired traction force generation in Vangl2Lp/+ TECs. YAP signaling in Vangl2Lp airway epithelium was reduced, consistent with a role for VANGL2 in mechanotransduction. Furthermore, activation of RhoA signaling restored actomyosin organization in Vangl2Lp/+, confirming RhoA as an effector of VANGL2. This study identifies a pivotal role for VANGL2 in mechanosignaling, which underlies the key role of the PCP pathway in tissue morphogenesis.
Dean C, Taylor B, Rice A, et al., 2020, Mechanism of lung development in the aetiology of adult congenital pulmonary airway malformations, Thorax, Vol: 75, Pages: 1001-1003, ISSN: 0040-6376
Congenital pulmonary airway malformations (CPAMs) are rare lung abnormalities that result in cyst formation and are associated with respiratory distress in infants and malignant potential in adults. The pathogenesis of CPAMs remains unknown but data suggest disruption of the normal proximo-distal programme of airway branching and differentiation. Here, we demonstrate that adult human CPAM are lined with epithelium that retains SOX-2 and thyroid transcription factor-1 immunohistochemical markers, characteristic of the developing lung. However, RALDH-1, another key marker, is absent. This suggests a more complex aetiology for CPAM than complete focal arrest of lung development and may provide insight to the associated risk of malignancy.
Singh S, Hind M, Jordan S, et al., 2020, Weaning by surgical tracheostomy and portable ventilators released ICU ventilators during coronavirus disease 2019 surge in London., Critical Care Explorations, Vol: 2, Pages: 1-3, ISSN: 2639-8028
RATIONALE: Poor lung health in adult life may occur partly through suboptimal growth and development, as suggested by epidemiological evidence pointing to early life risk factors. OBJECTIVES: To systematically investigate the effects of lung development genes on adult lung function. METHODS: Using UK Biobank data, we tested the association of 391 genes known to influence lung development with FVC and FEV1/FVC. We split the dataset into two random subsets of 207,616 and 138,411 individuals, using the larger to select the most promising signals and the smaller for replication. MEASUREMENTS AND MAIN RESULTS: We identified 55 genes, of which 36 (16 for FVC; 19 for FEV1/FVC; 1 for both) had not been identified in the largest, most recent genome-wide study of lung function. Most of these 36 signals were intronic variants; expression data from blood and lung tissue showed that the majority affect the expression of the genes they lie within. Further testing of 34 of these 36 signals in the CHARGE and SpiroMeta consortia showed that 16 replicated after Bonferroni correction and another 12 at nominal significance level. 53 of the 55 genes fell into four biological categories whose function is to regulate organ size and cell integrity (growth factors; transcriptional regulators; cell-cell adhesion; extra-cellular matrix), suggesting that these specific processes are important for adult lung health. CONCLUSIONS: Our study demonstrates the importance of lung development genes in regulating adult lung function and influencing both restrictive and obstructive patterns. Further investigation of these developmental pathways could lead to druggable targets.
Price S, Singh S, Ledot S, et al., 2020, Respiratory management in severe acute respiratory syndrome coronavirus 2 infection., European Heart Journal: Acute Cardiovascular Care, Vol: 9, Pages: 229-238, ISSN: 2048-8734
The severe acute respiratory syndrome coronavirus 2 pandemic is to date affecting more than a million of patients and is challenging healthcare professionals around the world. Coronavirus disease 2019 may present with a wide range of clinical spectrum and severity, including severe interstitial pneumonia with high prevalence of hypoxic respiratory failure requiring intensive care admission. There has been increasing sharing experience regarding the patient's clinical features over the last weeks which has underlined the need for general guidance on treatment strategies. We summarise the evidence existing in the literature of oxygen and positive pressure treatments in patients at different stages of respiratory failure and over the course of the disease, including environment and ethical issues related to the ongoing coronavirus disease 2019 infection.
Akram K, Yates L, Mongey R, et al., 2019, Time-lapse imaging of alveologenesis in mouse precision-cut lung slices, Bio-protocol, Vol: 9, ISSN: 2331-8325
Alveoli are the gas-exchange units of lung. The process of alveolar development,alveologenesis, is regulated by a complex network of signaling pathways that act on various cell typesincluding alveolar type I and II epithelial cells, fibroblasts and the vascular endothelium. Dysregulatedalveologenesis results in bronchopulmonary dysplasia in neonates and in adults, disrupted alveolarregeneration is associated with chronic lung diseases including COPD and pulmonary fibrosis.Therefore, visualizing alveologenesis is critical to understand lung homeostasis and for thedevelopment of effective therapies for incurable lung diseases. We have developed a technique tovisualize alveologenesis in real-time using a combination of widefield microscopy and imagedeconvolution of precision-cut lung slices. Here, we describe this live imaging technique in step-by-stepdetail. This time-lapse imaging technique can be used to capture the dynamics of individual cells withintissue slices over a long time period (up to 16 h), with minimal loss of fluorescence or cell toxicity.
Apps M, Pavitt M, Lewis A, et al., 2019, Increasing CPAP (Continuous Positive Airway Pressure) leads to increasing trans-pulmonary pressure with increased activity of the abdominal wall muscles to aid Expiration, International Congress of the European-Respiratory-Society (ERS), Publisher: EUROPEAN RESPIRATORY SOC JOURNALS LTD, ISSN: 0903-1936
Kim SY, Mongey R, Wang P, et al., 2019, LSC-2019-A novel ex-vivo approach to study lung injury and repair, European-Respiratory-Society (ERS) International Congress, Publisher: European Respiratory Society, ISSN: 0903-1936
Cockbain B, Price LC, Hind M, 2019, Bony breathlessness: reversible pulmonary hypertension in melnick-needles syndrome using noninvasive ventilation, Circulation, Vol: 140, Pages: 880-885, ISSN: 0009-7322
Patient presentation: A 48-year-old woman with Melnick-Needles Syndrome (MNS, a congenital bony dysplasia) was referred because ofprogressive breathlessness and exertional presyncope.She described 12 months of progressive exertional dyspnea with reduced exercise tolerance. She could walk just 10 m on the flat and foundher office-based job unmanageable. There were features of exertional presyncope but no collapse, chest pain, cough, or palpitations. She reportedan inability to sleep supine, nocturia, morning headaches, and poor-quality, unrefreshing sleep.She had snored since childhood, improving after micrognathia surgeryin her 20s. Smoking history was modest: a 3 pack-year exposure quitting adecade earlier. She consumed minimal alcohol and no recreational drugs.She took no regular medication and reported no recent foreign travel.Aside from MNS, she had no other notable history.
Ng-Blichfeldt J-P, Gosens R, Dean C, et al., 2019, Regenerative pharmacology for COPD: breathing new life into old lungs, Thorax, Vol: 74, Pages: 890-897, ISSN: 1468-3296
Chronic obstructive pulmonary disease (COPD) is a major global health concern with few effective treatments. Widespread destruction of alveolar tissue contributes to impaired gas exchange in severe COPD, and recent radiological evidence suggests that destruction of small airways is a major contributor to increased peripheral airway resistance in disease. This important finding might in part explain the failure of conventional anti-inflammatory treatments to restore lung function even in patients with mild disease. There is a clear need for alternative pharmacological strategies for patients with COPD/emphysema. Proposed regenerative strategies such as cell therapy and tissue engineering are hampered by poor availability of exogenous stem cells, discouraging trial results, and risks and cost associated with surgery. An alternative therapeutic approach is augmentation of lung regeneration and/or repair by biologically active factors, which have potential to be employed on a large scale. In favour of this strategy, the healthy adult lung is known to possess a remarkable endogenous regenerative capacity. Numerous preclinical studies have shown induction of regeneration in animal models of COPD/emphysema. Here, we argue that given the widespread and irreversible nature of COPD, serious consideration of regenerative pharmacology is necessary. However, for this approach to be feasible, a better understanding of the cell-specific molecular control of regeneration, the regenerative potential of the human lung and regenerative competencies of patients with COPD are required.
Liew F, Gargoum F, Potter R, et al., 2019, Platypnoea-orthodeoxia syndrome: beware of investigations undertaken supine, Thorax, Vol: 74, Pages: 917-919, ISSN: 1468-3296
Platypnoea-orthodeoxia syndrome (POS) is a rare disorder, manifesting as deoxygenation occurring when the patient is in the upright position. Four broad mechanisms for the condition have been described: intracardiac shunts, intrapulmonary shunts, hepatopulmonary syndrome and pulmonary ventilation-perfusion mismatch. Here, we present the first case of POS in a patient with a proven right to left intracardiac shunt occurring in the context of postural hypotension and normal right heart pressures. We highlight the need to carry out investigations in the upright position before discounting intracardiac shunting as a cause for the syndrome. Short-term improvement of the syndrome was obtained with medical management of the patient's orthostatic hypotension and as such suggests a conservative management strategy for similar patients, which may delay the need for invasive procedures to close the anatomical defect.
Taylor BA, Hind M, Rice A, et al., 2019, Investigating the Molecular Mechanisms of Congenital Pulmonary Airway Malformation (CPAM), 211th Meeting of the Pathological-Society-of-Great-Britain-and-Ireland, Publisher: WILEY, Pages: S11-S11, ISSN: 0022-3417
Akram K, Yates L, Mongey R, et al., 2019, Live imaging of alveologenesis in precision-cut lung slices reveals dynamic epithelial cell behaviour, Nature Communications, Vol: 10, Pages: 1-16, ISSN: 2041-1723
Damage to alveoli, the gas-exchanging region of the lungs, is a component of many chronic and acute lung diseases. In addition, insufficient generation of alveoli results in bronchopulmonary dysplasia, a disease of prematurity. Therefore visualising the process of alveolar development (alveologenesis) is critical for our understanding of lung homeostasis and for the development of treatments to repair and regenerate lung tissue. Using long-term, time-lapse imaging of precision-cut lung slices, we show alveologenesis for the first time. We reveal that during this process, epithelial cells are highly mobile and we identify specific cell behaviours that contribute to alveologenesis: cell clustering, hollowing and cell extension. Using the cytoskeleton inhibitors blebbistatin and cytochalasin D, we showed that cell migration is a key driver of alveologenesis. This study reveals important novel information about lung biology and provides a new system in which to manipulate alveologenesis genetically and pharmacologically.
Hind M, 2019, Chest pain, RESPIRATORY MEDICINE, 3RD EDITION, Editors: Palange, Rohde, Publisher: EUROPEAN RESPIRATORY SOCIETY, Pages: 108-109, ISBN: 978-1-84984-079-8
Proudfoot A, Bayliffe A, O'Kane CM, et al., 2018, Novel anti-tumour necrosis factor receptor-1 (TNFR1) domain antibody prevents pulmonary inflammation in experimental acute lung injury, Thorax, Vol: 73, Pages: 723-730, ISSN: 1468-3296
BACKGROUND: Tumour necrosis factor alpha (TNF-α) is a pleiotropic cytokine with both injurious and protective functions, which are thought to diverge at the level of its two cell surface receptors, TNFR1 and TNFR2. In the setting of acute injury, selective inhibition of TNFR1 is predicted to attenuate the cell death and inflammation associated with TNF-α, while sparing or potentiating the protective effects of TNFR2 signalling. We developed a potent and selective antagonist of TNFR1 (GSK1995057) using a novel domain antibody (dAb) therapeutic and assessed its efficacy in vitro, in vivo and in a clinical trial involving healthy human subjects. METHODS: We investigated the in vitro effects of GSK1995057 on human pulmonary microvascular endothelial cells (HMVEC-L) and then assessed the effects of pretreatment with nebulised GSK1995057 in a non-human primate model of acute lung injury. We then tested translation to humans by investigating the effects of a single nebulised dose of GSK1995057 in healthy humans (n=37) in a randomised controlled clinical trial in which subjects were subsequently exposed to inhaled endotoxin. RESULTS: Selective inhibition of TNFR1 signalling potently inhibited cytokine and neutrophil adhesion molecule expression in activated HMVEC-L monolayers in vitro (P<0.01 and P<0.001, respectively), and also significantly attenuated inflammation and signs of lung injury in non-human primates (P<0.01 in all cases). In a randomised, placebo-controlled trial of nebulised GSK1995057 in 37 healthy humans challenged with a low dose of inhaled endotoxin, treatment with GSK1995057 attenuated pulmonary neutrophilia, inflammatory cytokine release (P<0.01 in all cases) and signs of endothelial injury (P<0.05) in bronchoalveolar lavage and serum samples. CONCLUSION: These data support the potential for pulmonary delivery of a selective TNFR1 dAb as a novel therapeutic approach for the prevention of acute respiratory distress syndrome. TRI
Allen CJ, Freeman T, Perera W, et al., 2018, Thoracic Park: cardiac MRI reveals massive thoracic varices as consequence of inferior vena cava ligation, EHJ Cardiovascular Imaging / European Heart Journal - Cardiovascular Imaging, Vol: 19, Pages: 359-360, ISSN: 2047-2412
Alcada J, Shao D, Griffiths MJ, et al., 2018, Retinoic Acid Upregulates Endothelial-Derived Angiocrine Factors to Promote Human Alveolar Epithelial Repair, International Conference of the American-Thoracic-Society, Publisher: AMER THORACIC SOC, ISSN: 1073-449X
Pavitt MJ, Nevett J, Swanton LL, et al., 2017, London Ambulance source data on choking incidence for the calendar year 2016 – an observational study., BMJ Open Respiratory Research, Vol: 4, ISSN: 2052-4439
Introduction Complete foreign body airway obstruction is a life-threatening emergency, but there are limited data on its epidemiology.Methods We conducted a retrospective analysis of data collected routinely from London Ambulance Service calls coded as being for choking was undertaken for the calendar year of 2016.Results There were 1916 choking episodes of significant severity to call for emergency assessment in London during 2016, 0.2% of total calls requiring an ambulance response, an average of 5.2 per day. The incidence increased at the extremes of age. Calls coded as choking occurred at times consistent with lunch and dinner and less frequently at breakfast. Peak incidence occurred at Sunday lunchtimes and on Wednesday evenings.Conclusions Choking is a substantial health problem for Londoners to seek emergency assistance. Choking is more frequent at the extremes of age with a higher incidence at lunch and dinner time. Greater public awareness of choking and its management could help to prevent avoidable deaths.
Alcada J, Shao DS, Griffiths MJD, et al., 2017, MECHANISMS OF REGENERATION: RETINOIC ACID ACTS VIA THE ENDOTHELIUM TO DRIVE HUMAN LUNG REPAIR, Winter Meeting of the British-Thoracic-Society, Publisher: BMJ PUBLISHING GROUP, Pages: A106-A107, ISSN: 0040-6376
Pavitt M, Swanton L, Hind M, et al., 2017, Effectiveness of approaches to choking due to foreign body airway obstruction - a physiological study, European-Respiratory-Society (ERS) International Congress, Publisher: EUROPEAN RESPIRATORY SOC JOURNALS LTD, ISSN: 0903-1936
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