Distal Airways Research Articles

Biaxial mechanical properties of the bronchial tree: Characterization of elasticity, extensibility, and energetics, including the effect of strain rate and preconditioning

Distal airways commonly obstruct in lung disease and despite their importance, their mechanical properties are vastly underexplored. The lack of bronchial experiments restricts current airway models to either assume rigid structures, or extrapolate the material properties of the trachea to represent the small airways. Furthermore, past works are exclusively limited to uniaxial testing; investigating the multidirectional tensile loads of both the proximal and distal pulmonary airways is long overdue. Here we present comprehensive mechanical and viscoelastic properties of the porcine airway tree, including the trachea, trachealis muscle, large bronchi, and small bronchi, via measures of elasticity, extensibility, and energetics to explore regional and directional dependencies, cross-examining strain rate and preconditioning effects using planar equibiaxial tensile tests for the first time. We find bronchial regions are notably heterogeneous, where the trachea exhibits greater stiffness, energy loss, and preconditioning sensitivity than the smaller airways. Interestingly, the trachealis muscle is similar to the distal bronchi, despite being anatomically located adjacent to the proximal ring. Tissues are anisotropic and axially stiffer under initial loading, losing more energy with greater stress relaxation circumferentially. Strain rate dependency is also noted, where tissues are more energetically efficient at the faster strain rate, likely attributable to the microstructure. Findings highlight assumptions of homogeneity and isotropy are inadequate, and enable the improvement of aerosol flow and dynamic airway deformation computational predictive models. These results provide much needed fundamental material properties for future explorations contrasting healthy versus diseased pulmonary airway mechanics to better understand the relationship between structure and lung function. Statement of significanceWe present comprehensive multiaxial mechanical tensile experiments of the proximal and distal airways via measures of maximum stress, initial and ultimate moduli, strain and stress transitions, hysteresis, energy loss, and stress relaxation, and further assess preconditioning and strain rate dependencies to examine the relationship between lung function and structure. The mechanical response of the bronchial tree demonstrates significant anisotropy and heterogeneity, even within the tracheal ring, and emphasizes that contrary to past studies, the behavior of the proximal airways cannot be extended to distal bronchial tree analyses. Establishing these material properties is critical to advancing our understanding of airway function and in developing accurate computational simulations to help diagnose and monitor pulmonary diseases.

Population-wide gene disruption in the murine lung epithelium via AAV-mediated delivery of CRISPR-Cas9 components

With the aim of expediting drug target discovery and validation for respiratory diseases, we developed an optimized method for in situ somatic gene disruption in murine lung epithelial cells via AAV6-mediated CRISPR-Cas9 delivery. Efficient gene editing was observed in lung type II alveolar epithelial cells and distal airway cells following assessment of single- or dual-guide AAV vector formats, Cas9 variants, and a sequential dosing strategy with combinatorial guide RNA expression cassettes. In particular, we were able to demonstrate population-wide gene disruption within distinct epithelial cell types for separate targets in Cas9 transgenic animals, with minimal to no associated inflammation. We also observed and characterized AAV vector integration events that occurred within directed double-stranded DNA break sites in lung cells, highlighting a complicating factor with AAV-mediated delivery of DNA nucleases. Taken together, we demonstrate a uniquely effective approach for somatic engineering of the murine lung, which will greatly facilitate the modeling of disease and therapeutic intervention.

Effects of Aeroallergen Sensitization on Symptom Severity, Pulmonary Function, and Bronchodilator Response in Children With Bronchial Asthma

Background: Allergen sensitization, symptom severity, pulmonary function test, and bronchodilator response are important in the diagnosis and treatment of asthma. However, the relationship between these factors remains unclear. Objective: The objective of this study was to investigate the relationship between aeroallergen sensitization and asthma severity, pulmonary function, and bronchodilator response among pediatric patients with bronchial asthma. Methods: This was a prospective study where 155 pediatric patients aged 7–18 years old with bronchial asthma were recruited from Outpatient Clinics. Patients who met the inclusion criteria proceeded with spirometry and aeroallergen skin prick test. Results: There was a significant degree of sensitization, wherein 100% of the patients had sensitization to one or more aeroallergens. Among these children, 106 (68%) were polysensitized. The polysensitized group had more severe and persistent asthma severity profile (p<0.001) and worse pulmonary function (p<0.001). The frequency of abnormal pre-bronchodilator lung function of the polysensitized group was higher than the monosensitized group (p<0.001). A positive bronchodilator response was higher among polysensitized children as compared to monosensitized children (p<0.001). Sensitization to Dermatophagoides farinae, Dermatophagoides pteronyssinus and dog correlated with impairment of both the large airways and distal small airways while sensitization to cat, cockroach, and horse correlated only with impairment of the large airways (p<0.05). Patients sensitized to D. farinae, D. pteronyssinus, cat and dog had significant bronchodilator response (p<0.05). Conclusion: Polysensitized asthmatic children had a more persistent and severe asthma profile, worse pulmonary function, and higher bronchodilator reversibility compared to the monosensitized group.

IPSC-Derived Airway Epithelial Cells: Progress, Promise, and Challenges.

Induced pluripotent stem cells (iPSCs) generated from somatic cell sources are pluripotent and capable of indefinite expansion in vitro. They provide an unlimited source of cells that can be differentiated into lung progenitor cells for potential clinical use in pulmonary regenerative medicine. This review gives a comprehensive overview of recent progress toward the use of iPSCs to generate proximal and distal airway epithelial cells and mix lung organoids. Furthermore, their potential applications and future challenges for the field are discussed, with a focus on the technological hurdles that must be cleared before stem cell therapeutics can be used for clinical treatment.

CRYPTOGENIC ORGANIZING PNEUMONIA: A DIAGNOSTIC DILEMMA IN A WORLD DOMINATED BY COVID-19 PNEUMONIA

CRYPTOGENIC ORGANIZING PNEUMONIA: A DIAGNOSTIC DILEMMA IN A WORLD DOMINATED BY COVID-19 PNEUMONIA

A CASE OF SECONDARY ORGANIZING PNEUMONIA FROM INFLIXIMAB

A CASE OF SECONDARY ORGANIZING PNEUMONIA FROM INFLIXIMAB

SEALANT GLUE TO MANAGE BLEEDING POSTNAVIGATION BRONCHOSCOPY-GUIDED LUNG BIOPSY

SEALANT GLUE TO MANAGE BLEEDING POSTNAVIGATION BRONCHOSCOPY-GUIDED LUNG BIOPSY

RECANNULATION OF COMPLETELY OCCLUDED BRONCHUS INTERMEDIUS WITH A DIODE LASER

RECANNULATION OF COMPLETELY OCCLUDED BRONCHUS INTERMEDIUS WITH A DIODE LASER

DIAGNOSTIC ACCURACY OF ROBOTIC-ASSISTED BRONCHOSCOPY USING SHAPE-SENSING TECHNOLOGY WITH CONE-BEAM CT IMAGING IN EVALUATION OF PULMONARY LESIONS: A RETROSPECTIVE REVIEW

DIAGNOSTIC ACCURACY OF ROBOTIC-ASSISTED BRONCHOSCOPY USING SHAPE-SENSING TECHNOLOGY WITH CONE-BEAM CT IMAGING IN EVALUATION OF PULMONARY LESIONS: A RETROSPECTIVE REVIEW

POST-TRANSPLANT BRONCHIAL STENOSIS: A SINGLE CENTER RETROSPECTIVE STUDY

POST-TRANSPLANT BRONCHIAL STENOSIS: A SINGLE CENTER RETROSPECTIVE STUDY

ROLE OF VENOVENOUS EXTRACORPOREAL MEMBRANE OXYGENATION IN SALVAGE THERAPY FOR CENTRAL AIRWAY OBSTRUCTION: A CASE REPORT

ROLE OF VENOVENOUS EXTRACORPOREAL MEMBRANE OXYGENATION IN SALVAGE THERAPY FOR CENTRAL AIRWAY OBSTRUCTION: A CASE REPORT

TRACHEAL AND BRONCHIAL STENOSIS CAUSED BY CHRONIC UNCONTROLLED GASTROESOPHAGEAL REFLUX DISEASE

TRACHEAL AND BRONCHIAL STENOSIS CAUSED BY CHRONIC UNCONTROLLED GASTROESOPHAGEAL REFLUX DISEASE

BILATERAL PNEUMOTHORAX, PNEUMOMEDIASTINUM, AND SUBCUTANEOUS EMPHYSEMA: A CASE OF A RARE SYNCHRONOUS TRIAD IN ASTHMA EXACERBATION

BILATERAL PNEUMOTHORAX, PNEUMOMEDIASTINUM, AND SUBCUTANEOUS EMPHYSEMA: A CASE OF A RARE SYNCHRONOUS TRIAD IN ASTHMA EXACERBATION

Adeno-associated virus mediates gene transduction after static cold storage treatment in rodent lung transplantation

ObjectiveAdeno-associated virus is a clinically used gene therapy vector but has not been studied in lung transplantation. We sought to determine the efficacy of adeno-associated virus delivery during static cold storage via the airway versus the pulmonary artery before lung transplantation in a rodent model. MethodsLewis rat lung grafts were treated with a dose of 8e8 or 4e9 viral genome/μL recombinant adeno-associated virus subtype-9 vectors containing firefly luciferase genomes administered via the pulmonary artery or airway during cold storage. A control group did not receive adeno-associated virus. Recipient syngeneic rats then underwent single left lung transplantation. Animals underwent bioluminescence imaging on postoperative days 7, 14, 28, and 56. Explanted tissues were prepared as lysates to quantify luciferase activity. Immunohistochemistry was performed to evaluate cellular transgene expression patterns. ResultsControl animals with no luminescent signal produced a background radiance of 6.1e4 p/s/cm2/sr. In the airway delivery group, mean radiance was greater than the control at 4e9 viral genome/μL postoperative day 7 radiance 6.9e4 p/s/cm2/sr (P = .04). In the pulmonary artery delivery group, we observed greater in vivo luminescence in animals receiving 4e9 viral genome/μL compared with all other groups. However, analysis of tissue lysate revealed greater luminescence in the airway delivery group and suggested off-target expression in heart and liver tissue in the pulmonary artery delivery group. Immunohistochemistry demonstrated transgene staining in distal airway epithelium and alveoli but sparing of the vasculature in the airway delivery group. ConclusionsAdeno-associated virus mediates gene transduction during static cold storage in rat lung isografts when administered via the airway and pulmonary artery. Airway administration leads to robust transgene expression in respiratory epithelial cells, whereas pulmonary artery administration targets alternative cell types and increases extrapulmonary transgene expression.

Cytokines Profile and Lung Function in Children with Obesity and Asthma: A Case Control Study.

The existence of common inflammatory biomarkers linking obesity and asthma in children has been hypothesized. Nevertheless, laboratory and clinical characteristics of children with obesity and asthma are still poorly defined. The primary aim of the present study is to investigate the lung function and the cytokine profile, in children with obesity and asthma. In this prospective, cross-sectional pilot study, pulmonary function tests, biochemical parameters, and serum cytokines levels were compared in three groups of 28 children each, matched for age and sex. Obese children showed normal forced spirometry values except an increased distal airway resistance in subjects with obesity and no asthma. Both groups including obese children showed higher leptin and IL-10 levels and lower adiponectin and TNF-alpha levels compared to children with no obesity and asthma. IL-33 and TGF-beta1 levels were higher in children with obesity and asthma vs. children with normal weight and asthma. Finally, IL-6 was undetectable in approximately 70% of obese children with no asthma, in 57% obese asthmatic children and in 100% of children with normal-weight and asthma. Children with obesity and asthma show the most striking cytokine profile, suggesting a pro-inflammatory role of fat mass in asthma development.

Epithelial Endoplasmic Reticulum Stress Enhances the Risk of Muc5b-associated Lung Fibrosis.

The gain-of-function minor allele of the MUC5B (mucin 5B, oligomeric mucus/gel-forming) promoter (rs35705950) is the strongest risk factor for idiopathic pulmonary fibrosis (IPF), a devastating fibrotic lung disease that leads to progressive respiratory failure in adults. We have previously demonstrated that Muc5b overexpression in mice worsens lung fibrosis after bleomycin exposure and have hypothesized that excess Muc5b promotes endoplasmic reticulum (ER) stress and apoptosis, stimulating fibrotic lung injury. Here, we report that ER stress pathway members ATF4 (activating transcription factor 4) and ATF6 coexpress with MUC5B in epithelia of the distal IPF airway and honeycomb cyst and that this is more pronounced in carriers of the gain-of-function MUC5B promoter variant. Similarly, in mice exposed to bleomycin, Muc5b expression is temporally associated with markers of ER stress. Using bulk and single-cell RNA sequencing in bleomycin-exposed mice, we found that pathologic ER stress-associated transcripts Atf4 and Ddit3 (DNA damage inducible transcript 3) were elevated in alveolar epithelia of SFTPC-Muc5b transgenic (SFTPC-Muc5bTg) mice relative to wild-type (WT) mice. Activation of the ER stress response inhibits protein translation for most genes by phosphorylation of Eif2α (eukaryotic translation initiation factor 2 alpha), which prevents guanine exchange by Eif2B and facilitates translation of Atf4. The integrated stress response inhibitor (ISRIB) facilitates interaction of phosphorylated Eif2α with Eif2B, overcoming translation inhibition associated with ER stress and reducing Atf4. We found that a single dose of ISRIB diminished Atf4 translation in SFTPC-Muc5bTg mice after bleomycin injury. Moreover, ISRIB resolved the exaggerated fibrotic response of SFTPC-Muc5bTg mice to bleomycin. In summary, we demonstrate that MUC5B and Muc5b expression is associated with pathologic ER stress and that restoration of normal translation with a single dose of ISRIB promotes lung repair in bleomycin-injured Muc5b-overexpressing mice.

Predicting asthma exacerbations: is there utility in noninvasive assessment of distal airway inflammation using multiple flowFENO?

<b>Alveolar inflammation, as assessed by partitioned exhaled nitric oxide, appears to provide promising insights into small airway disease relevant to asthma exacerbation</b>https://bit.ly/38KqyCk

Iraq/Afghanistan war lung injury reflects burn pits exposure

This descriptive case series retrospectively reviewed medical records from thirty-one previously healthy, war-fighting veterans who self-reported exposure to airborne hazards while serving in Iraq and Afghanistan between 2003 and the present. They all noted new-onset dyspnea, which began during deployment or as a military contractor. Twenty-one subjects underwent non-invasive pulmonary diagnostic testing, including maximum expiratory pressure (MEP) and impulse oscillometry (IOS). In addition, five soldiers received a lung biopsy; tissue results were compared to a previously published sample from a soldier in our Iraq Afghanistan War Lung Injury database and others in our database with similar exposures, including burn pits. We also reviewed civilian control samples (5) from the Stony Brook University database. Military personnel were referred to our International Center of Excellence in Deployment Health and Medical Geosciences, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell under the auspices of Northwell IRB: 17-0140-FIMR Feinstein Institution for Medical Research “Clinicopathologic characteristics of Iraq Afghanistan War Lung Injury.” We retrospectively examined medical records, including exposure data, radiologic imaging, and non-invasive pulmonary function testing (MGC Diagnostic Platinum Elite Plethysmograph) using the American Thoracic Society (ATS) standard interpretation based on Morgan et al., and for a limited cohort, biopsy data. Lung tissue, when available, was examined for carbonaceous particles, polycyclic aromatic hydrocarbons (Raman spectroscopy), metals, titanium connected to iron (Brookhaven National Laboratory, National Synchrotron Light Source II, Beamline 5-ID), oxidized metals, combustion temperature, inflammatory cell accumulation and fibrosis, neutrophil extracellular traps, Sirius red, Prussian Blue, as well as polarizable crystals/particulate matter/dust. Among twenty-one previously healthy, deployable soldiers with non-invasive pulmonary diagnostic tests, post-deployment, all had severely decreased MEP values, averaging 42% predicted. These same patients concurrently demonstrated abnormal airways reactance (X5Hz) and peripheral/distal airways resistance (D5–D20%) via IOS, averaging − 1369% and 23% predicted, respectively. These tests support the concept of airways hyperresponsiveness and distal airways narrowing, respectively. Among the five soldiers biopsied, all had constrictive bronchiolitis or bronchiolitis or severe pulmonary fibrosis. We detected the presence of polycyclic aromatic hydrocarbons (PAH)—which are products of incomplete combustion—in the lung tissue of all five warfighters. All also had detectable titanium and iron in the lungs. Metals were all oxidized, supporting the concept of inhaling burned metals. Combustion temperature was consistent with that of burned petrol rather than higher temperatures noted with cigarettes. All were nonsmokers. Neutrophil extracellular traps were reported in two biopsies. Compared to our prior biopsies in our Middle East deployment database, these histopathologic results are similar, since all database biopsies have constrictive bronchiolitis, one has lung fibrosis with titanium bound to iron in fixed mathematical ratios of 1:7 and demonstrated polarizable crystals. These results, particularly constrictive bronchiolitis and polarizable crystals, support the prior data of King et al. (N. Engl. J. Med. 365:222–230, 2011) Soldiers in this cohort deployed to Iraq and Afghanistan since 2003, with exposure to airborne hazards, including sandstorms, burn pits, and improvised explosive devices, are at high risk for developing chronic clinical respiratory problems, including: (1) reduction in respiratory muscle strength; (2) airways hyperresponsiveness; and (3) distal airway narrowing, which may be associated with histopathologic evidence of lung damage, reflecting inhalation of burned particles from burn pits along with particulate matter/dust. Non-invasive pulmonary diagnostic tests are a predictor of burn pit-induced lung injury.

Proteomic Profiling of Hypoplastic Lungs Suggests an Underlying Inflammatory Response in the Pathogenesis of Abnormal Lung Development in Congenital Diaphragmatic Hernia.

The pathogenesis of lung hypoplasia in congenital diaphragmatic hernia (CDH), a common birth defect, is poorly understood. The diaphragmatic defect can be repaired surgically, but the abnormal lung development contributes to a high mortality in these patients. To understand the underlying pathobiology, we compared the proteomic profiles of fetal rat lungs at the alveolar stage (E21) that were either exposed to nitrofen in utero (CDH lungs, n=5) or exposed to vehicle only (non-CDH control lungs, n=5). Pathway analysis of proteomic datasets showed significant enrichment in inflammatory response proteins associated with cytokine signaling and Epstein Barr Virus in nitrofen CDH lungs. Among the 218 significantly altered proteins between CDH and non-CDH control lungs were Tenascin C, CREBBP, LYN, and STAT3. We showed that Tenascin C was decreased around the distal airway branches in nitrofen rat lungs and human CDH lungs, obtained from stillborn fetuses that did not receive pre- or postnatal treatment. In contrast, STAT3 was significantly increased in the airway epithelium of nitrofen lungs at E21. STAT3 inhibition after direct nitrofen exposure to fetal rat lung explants (E14.5) partially rescued the hypoplastic lung phenotype ex vivo by increasing peripheral lung budding. Moreover, we demonstrated that several STAT3-associated cytokines (IL-15, IL-9, andIL-2) are increased in fetal tracheal aspirates of CDH survivors compared with nonsurvivors after fetoscopic endoluminal tracheal occlusion. With our unbiased proteomics approach, we showed for the first time that downstream inflammatory processes are likely involved in the pathogenesis of abnormal lung development in CDH.

Suitability of Bronchoscopic Spraying for Fluid Deposition in Lower Airway Regions: Fluorescence Analysis on a Transparent In Vitro Airway Model.

Introduction: Bronchoscopic spraying has potential for the application of therapeutic drugs in distal regions of the lung by bypassing the upper airways. However, there is a lack of understanding about the underlying fluid transport phenomena that are responsible for the intrapulmonary propagation of applied liquid. Methods: By using a transparent airway model, this study provides first experimental insights into relevant transport phenomena of bronchoscopic spraying. Furthermore, the penetration depth of the application is quantitatively evaluated. Laser-induced fluorescence is used to analyze fluid propagation in the transparent channels. Potential influencing factors such as the positioning in different airways, application number, breathing pattern, and lung obstructions are varied within this study to determine their influence on liquid deposition. Findings: This study shows that the method of bronchoscopic spraying allows the application of liquid in distal regions of the airway model. The position of the bronchoscope is a key influencing factor in increasing the penetration depth. We found that fluid transport along the distal airways essentially occurs by the film and plug flow phenomenon during application, which is similar to the transport mechanisms during instillation. Liquid plugs in lower airways are responsible for the reorganization of liquid during proximal movements and thereby influence the penetration depth in subsequent applications.