The role of nasal congestion as a defence against respiratory viruses.

INTRODUCTION: This review discusses how nasal congestion may have benefits as a mechanism of defence against respiratory viruses. METHODS: A literature research was conducted on respiratory viruses and nasal congestion, following a recently published review on how temperature sensitivity is important for the success of common respiratory viruses. RESULTS: The literature reported that common respiratory viruses are temperature sensitive and replicate well at the cooler temperatures of the upper airways (32°C), but replication is restricted at body temperature (37°C). The amplitude of the phases of congestion and decongestion associated with the nasal cycle was increased on infection with respiratory viruses and this caused unilateral nasal congestion and obstruction. Nasal congestion and obstruction increase nasal mucosal temperature towards 37°C and therefore restricted the replication of respiratory viruses. CONCLUSION: Nasal congestion associated with the nasal cycle may act as a mechanism of respiratory defence against infection with respiratory viruses.

Functional analysis and evaluation of respiratory cilia in healthy Chinese children.

BACKGROUND: To aid in the diagnosis of Primary Ciliary Dyskinesia (PCD) and to evaluate the respiratory epithelium in respiratory disease, normal age-related reference ranges are needed for ciliary beat frequency (CBF), beat pattern and ultrastructure. Our aim was to establish reference ranges for healthy Chinese children. METHODS: Ciliated epithelial samples were obtained from 135 healthy Chinese children aged below 18 years by brushing the inferior nasal turbinate. CBF and beat pattern were analysed from high speed video recordings. Epithelial integrity and ciliary ultrastructure were assessed using transmission electronic microscopy. RESULTS: The mean CBF from 135 children studied was 10.1 Hz (95% CI 9.8 to 10.4). Approximately 20% (ranged 18.0-24.2%) of ciliated epithelial edges were found to have areas of dyskinetically beating cilia. Normal beat pattern was observed in ciliated epithelium from all subjects. We did not find any effect of exposure to second hand smoke on CBF in our subjects. Microtubular defects were found in 9.3% of all of the cilia counted in these children, while other ciliary ultrastructural defects were found in less than 3%. CONCLUSIONS: We established the reference range for CBF, beat pattern and ultrastructure in healthy Chinese children. Using similar methodology, we found a lower overall mean CBF than previously obtained European values. This study highlights the need to establish normative data for ciliary function in different populations.

Nasal-associated lymphoid tissues (NALTs) support the recall but not priming of influenza virus-specific cytotoxic T cells.

The lymphoid tissue that drains the upper respiratory tract represents an important induction site for cytotoxic T lymphocyte (CTL) immunity to airborne pathogens and intranasal vaccines. Here, we investigated the role of the nasal-associated lymphoid tissues (NALTs), which are mucosal-associated lymphoid organs embedded in the submucosa of the nasal passage, in the initial priming and recall expansion of CD8+ T cells following an upper respiratory tract infection with a pathogenic influenza virus and immunization with a live attenuated influenza virus vaccine. Whereas NALTs served as the induction site for the recall expansion of memory CD8+ T cells following influenza virus infection or vaccination, they failed to support activation of naïve CD8+ T cells. Strikingly, NALTs, unlike other lymphoid tissues, were not routinely surveyed during the steady state by circulating T cells. The selective recruitment of memory T cells into these lymphoid structures occurred in response to infection-induced elevation of the chemokine CXCL10, which attracted CXCR3+ memory CD8+ T cells. These results have significant implications for intranasal vaccines, which deliver antigen to mucosal-associated lymphoid tissue and aim to elicit protective CTL-mediated immunity.

Measurement of mucociliary clearance in the patients with multiple sclerosis.

OBJECTIVES: The aim of the present study was to measure nasal mucociliary clearance (NMC) time in the patients with MS and to compare the findings with healthy population. METHODS: Totally 97 individuals including 47 patients with relapsing-remitting multiple sclerosis and 50 healthy volunteers were enrolled into the study. Saccharin clearance test was performed on both groups and NMC time was measured. Data analysis was performed by SPSS version 24.0 statistics program (SPSS Inc., Chicago, Illinois, USA). Statistical tests were interpreted at p < 0.05 significance level. RESULTS: The NMC time averages in MS patients and healthy control group were 12.43 ± 4.05 min and 8.14 ± 2.87 min, respectively; the difference between the groups was significant (p < 0.001). There was a statistically strong association between NMC time values and Expanded Disability Status Scale (EDSS) values in MS patients (r = 0.817, p < 0.001). CONCLUSION: We found nasal mucociliary transport time longer in MS patients than healthy population in the present study. To the best of our knowledge, there is not any study conducted about this topic in the literature. We believe that our findings would shed a light on further studies.

Amygdala-stimulation-induced apnea is attention and nasal-breathing dependent.

OBJECTIVE: Evidence suggests that disordered breathing is critically involved in Sudden Unexpected Death in Epilepsy (SUDEP). To that end, evaluating structures that are activated by seizures and can activate brain regions that produce cardiorespiratory changes can further our understanding of the pathophysiology of SUDEP. Past preclinical studies have shown that electrical stimulation of the human amygdala induces apnea, suggesting a role for the amygdala in controlling respiration. In this study, we aimed to both confirm these findings in a larger group of patients with intractable temporal lobe epilepsy (TLE) and also further explore the anatomical and cognitive properties of this effect. METHODS: Seven surgical TLE patients had depth electrodes implanted in the amygdala that were used to deliver electrical stimulation during functional mapping preceding resection. Real-time respiratory monitoring was performed in each patient to confirm apnea. RESULTS: Our data confirm that amygdala stimulation reliably induces apnea (occurring in all 7 patients) and further suggest that apnea can be overcome by instructing the patient to inhale, and can be prevented entirely by breathing through the mouth before electrical stimulation. Finally, stimulation-induced apnea occurred only when stimulating the medial-most amygdalar contacts located in the central nucleus. INTERPRETATION: These findings confirm a functional connection between the amygdala and respiratory control in humans. Moreover, they suggest specific amygdalar nuclei may be critical in mediating this effect and that attentional state is critical to apnea mediated by amygdala activation-perhaps alluding to future development of strategies for the prevention of SUDEP. Ann Neurol 2018;83:460-471.

TGF-ß1 Induces Epithelial-Mesenchymal Transition of Chronic Sinusitis with Nasal Polyps through MicroRNA-21.

OBJECTIVES: To characterize the epithelial-mesenchymal transition (EMT) in chronic rhinosinusitis with nasal polyps (CRSwNP) and to investigate the mechanism by which microRNA-21 (miR-21) regulates EMT in CRSwNP. METHOD: (1) Tissue experiments: Mucosa tissues were collected from 13 patients with CRSwNP and 12 patients with CRS without nasal polyps (CRSsNP), as well as 11 patients without CRS (controls). Protein localization and quantification were achieved by immunofluorescence staining and Western blotting, involving the epithelial marker protein E-cadherin and the mesenchymal marker proteins α-smooth muscle actin (α-SMA), fibronectin, and vimentin. Quantitative RT-PCR was used to detect the relative expression levels of miR-21 and TGF-ß1 mRNAs. (2) Cellular experiments: Primary human nasal epithelial cells (PHNECs) treated with TGF-ß1, or TGF-ß1 with miR-21 inhibitor, or miR-21 mimics alone were observed for morphology changes under a phase-contrast microscope. The expression levels of epithelial/mesenchymal marker proteins were determined as aforementioned. PTEN and phosphorylated Akt were detected by Western blotting. RESULTS: (1) Tissue experiments: Compared with the CRSsNP and control groups, the expression of E-cadherin was downregulated in the CRSwNP group, whereas the expression of TGF-ß1, α-SMA, fibronectin, and vimentin was upregulated. The expression levels of miR-21 and TGF-ß1 mRNAs in CRSwNP were significantly higher than those in CRSsNP and controls. (2) Cellular experiments: TGF-ß1 induced EMT-like transformation in PHNECs, featured by changes in cell morphology and upregulation of mesenchymal proteins and miR-21. The miR-21 inhibitor, as well as the Akt-specific -inhibitor, suppressed TGF-ß1-induced EMT. Mechanically, downregulation of miR-21 resulted in increased PTEN and decreased Akt phosphorylation. Furthermore, overexpression of miR-21 had the opposite effects. CONCLUSIONS: Our findings suggest that the TGF-ß1-miR-21-PTEN-Akt axis may contribute to the pathogenesis of CRSwNP. miR-21 might be a reliable target for treating nasal polyp genesis through EMT suppression. Moreover, miR-21 inhibitors could be a novel class of antipolyp drug that modulates PTEN expression and Akt activation. In addition, further investigation regarding the reason underlying miR-21 overexpression in CRSwNP could provide a molecular target for novel treatment strategies for nasal polyposis.

Nasal Cavity of Green Sea Turtles Contains 3 Independent Sensory Epithelia.

The morphological and histological features of the nasal cavity are diverse among animal species, and the nasal cavities of terrestrial and semiaquatic turtles possess 2 regions lined with each different type of sensory epithelium. Sea turtles can inhale both of volatile and water-soluble odorants with high sensitivity, but details of the architectural features and the distribution of the sensory epithelia within the sea turtle nasal cavity remain uncertain. The present study analyzed the nasal cavity of green sea turtles using morphological, computed tomographic, and histological methods. We found that the middle region of the sea turtle nasal cavity is divided into anterodorsal, anteroventral, and posterodorsal diverticula and a posteroventral excavation by connective tissue containing cartilages. The posterodorsal diverticulum was lined with a thin sensory epithelium, and the anterodorsal and anteroventral diverticula were occupied by a single thick sensory epithelium. In addition, a relatively small area on the posteroventral excavation was covered by independent sensory epithelium that differed from other 2 types of epithelia, and a single thin bundle derived from the posteroventral excavation comprised the most medial nerve that joins the anterior end of the olfactory nerve tract. These findings suggested that the posteroventral excavation identified herein transfers stimuli through an independent circuit and plays different roles when odorants arise from other nasal regions.

Sinus floor elevation in sites with a perforated schneiderian membrane: What is the effect of placing a collagen membrane in a rabbit model?

OBJECTIVE: To investigate the healing following sinus grafting in sites with a perforated schneiderian membrane repaired using a collagen membrane, compared to control sites without membrane perforation. MATERIALS & METHODS: Following elevation of the sinus membrane in 16 rabbits, each sinus was assigned to one of the following groups: (a) intentional schneiderian membrane perforation, followed by the placement of a collagen membrane and bone grafting (group SMP) and (b) bone grafting without a perforation of the schneiderian membrane and without a collagen membrane placement (control group). At 2 and 4 weeks (n = 8 for each time-point), microcomputed tomographic (micro-CT) and histomorphometric analyses were performed. RESULTS: Overall new bone formation in group SMP was significantly delayed compared to the control group at 2 and 4 weeks (1.58 ± 1.25% vs. 9.23 ± 2.69% at 2 weeks, 10.43 ± 3.55 vs. 17.86 ± 4.11% at 4 weeks, p < 0.05). At 2 weeks, new bone formation for the areas close to lateral (1.19 ± 2.02%) and medial sinus bone walls (3.17 ± 1.98%) was markedly delayed in group SMP compared to the control group (13.08 ± 6.13% and 12.75 ± 5.63%, respectively, p < 0.05), but there was no statistical difference in those areas at 4 weeks (p > 0.05). The augmented volumes at 2 and 4 weeks were not statistically significantly different in both groups. CONCLUSION: The perforation of the schneiderian membrane and the repair using a collagen membrane delayed new bone formation in the augmented sinuses. However, the extension of the collagen membrane on the sinus bone walls was also attributable to this delayed bone formation.

Effect of the temperature of nasal lavages on mucociliary clearance: a randomised controlled trial.

BACKGROUND: The respiratory epithelium is mainly constituted by caliciform (produces mucus, responsible of keeping moisture and trapping particles) and ciliated cells (transports mucus into the pharynx, by the movement of multiple cilia). For centuries, nasal lavages have been used for different rhinosinusal conditions. Some studies suggest not only a direct effect on the mobilisation of secretions, but also an improvement in mucociliary clearance rates. To our knowledge, the impact of temperature in nasal lavages has been scarcely studied. METHODOLOGY/PRINCIPAL: We used the saccharin test-applying it in the inferior turbinate and timing the detection of its taste-, to estimate mucociliary clearance rates before and after nasal lavages with saline solution at room (20 °C) or body (37 °C) temperatures. RESULTS: 78 healthy subjects were studied, with a mean saccharin test time of 13.88 min. Then, a nasal lavage was performed, half with 20 °C saline and the other with 37 °C. In both, times improved from baseline (from 13.66 to 11.59 and 14.06 to 9.4 min, respectively) with p values < 0.05. CONCLUSIONS: Nasal lavages with saline solution improve mucociliary clearance as measured by saccharin test. Temperature seems to matter, which should be taken into account when indicating nasal lavages to our patients.

Mechanical characteristics of the maxillary sinus Schneiderian membrane ex vivo.

OBJECTIVES: It has been speculated that certain Schneiderian membrane thickness (SMT) might be more prone to perforation. This investigation was aimed at studying the mechanical characteristics of the Schneiderian membrane under one- and two-dimensional tests and their correlation to the histological SMT in human samples. MATERIAL AND METHODS: Sixteen Schneiderian membranes were collected from 11 cadaver heads treated with Thiel's embalming method. The samples were processed and analyzed clinically and histologically. One-dimensional maximum elongation until perforation and two-dimensional resistance to ball penetration were performed after the biopsy. Data was analyzed by using the Wilcoxon rank test and the Spearman's rank correlation. RESULTS: The histological SMT was 1.36 ± 0.42 mm, whereas the clinical thickness was 0.27 ± 0.21 mm, yielding statistical significance (p = 0.000). The resistance under ball penetration was 0.59 ± 0.43 N and the mean maximum elongation in the one-dimension test 11.19 ± 7.14 mm. Expressed in percentage, the mean stretch was 241.36 ± 227.97% (range 31.5 up to 947%). A weak positive correlation was found between the ball penetration test and the SMT (r = 0.10, p = 0.711), while a weak negative correlation was found between stretching test and the SMT (r = -0.021, p = 0.94). CONCLUSIONS: Mechanical tests seem to indicate that SMT might not significantly predispose to Schneiderian membrane perforation. Hence, other anatomical and operator's factors should be considered of surpassing importance. CLINICAL RELEVANCE: Thinner SM might be more prone to perforation when detaching it from the maxillary sinus antrum; however, a thick membrane is not prevented to tear, as their resistance under elastic forces is not higher than thinner ones.

Enhanced chemosensory sensitivity in patients with idiopathic rhinitis and its reversal by nasal capsaicin treatment.

BACKGROUND: The therapeutic action of capsaicin treatment in patients with idiopathic rhinitis (IR) is based on ablation of the transient receptor potential cation channel subfamily V, receptor 1 (TRPV1)-substance P nociceptive signaling pathway. However, the functional consequences of capsaicin treatment on nasal nerve activation and the association between the reduction in nasal hyperreactivity (NHR) and response to capsaicin treatment remain unknown. OBJECTIVE: We sought to study the effects of capsaicin nasal spray on the afferent innervation of the nasal mucosa by monitoring trigeminal nerve activity in patients with IR and healthy control (HC) subjects. METHODS: A double-blind, placebo-controlled randomized trial with capsaicin nasal spray was performed involving 33 patients with IR and 12 HC subjects. Before and at 4, 12, and 26 weeks after treatment, nasal mucosal potentials (NMPs) were measured while exposing the nasal mucosa of patients with IR and HC subjects to aerosols with increasing doses of the chemical irritants allyl isothiocyanate (AITC; also known as mustard oil) or capsaicin. The threshold for each compound was determined for each subject. The results of the NMP measurements were evaluated in parallel with the therapeutic response, visual analog scale scores for nasal symptoms, self-reported NHR, and mRNA expression of PGP9.5; TRPV1; transient receptor potential cation channel subfamily A, receptor 1 (TRPA1); TRPV4; transient receptor potential cation channel subfamily M, member 8 (TRPM8); and nerve growth factor (NGF) in nasal biopsy specimens. RESULTS: AITC turned out to be the best stimulus because the coughing induced by capsaicin interfered with measurements. At baseline, the threshold for evoking changes in NMPs based on AITC was significantly lower for patients with IR compared with HC subjects (P = .0423). Capsaicin treatment of IR patients increased the threshold for the response to AITC at 4 and 12 weeks compared with placebo (P = .0406 and P = .0325, respectively), which returned to baseline by week 26 (P = .0611). This increase correlated with changes in visual analog scale major symptom (P = .0004) and total symptom (P = .0018) scores. IR patients with self-reported NHR at baseline showed a trend to being better responders to capsaicin treatment compared with patients with IR but without NHR (P = .10). CONCLUSION: The lower threshold for AITC based on NMPs in patients with IR compared with HC subjects and the increased threshold for AITC after capsaicin treatment in patients with IR demonstrate the crucial role of TRPA1 and TRPV1 in IR pathophysiology. The strong correlation between the increase in AITC threshold in patients with IR and symptom reduction after capsaicin treatment demonstrates the clinical relevance of these findings.

Novel applications for a noninvasive sampling method of the nasal mucosa.

Reliable methods for sampling the nasal mucosa provide clinical researchers with key information regarding respiratory biomarkers of exposure and disease. For quick and noninvasive sampling of the nasal mucosa, nasal lavage (NL) collection has been widely used as a clinical tool; however, limitations including volume variability, sample dilution, and storage prevent NL collection from being used in nonlaboratory settings and analysis of low abundance biomarkers. In this study, we optimize and validate a novel methodology using absorbent Leukosorb paper cut to fit the nasal passage to extract epithelial lining fluid (ELF) from the nasal mucosa. The ELF sampling method limits the dilution of soluble mediators, allowing quantification of both high- and low-abundance soluble biomarkers such as IL-1ß, IL-8, IL-6, interferon gamma-induced protein 10 (IP-10), and neutrophil elastase. Additionally, we demonstrate that this method can successfully detect the presence of respiratory pathogens such as influenza virus and markers of antibiotic-resistant bacteria in the nasal mucosa. Efficacy of ELF collection by this method is not diminished in consecutive-day sampling, and percent recovery of both recombinant IL-8 and soluble mediators are not changed despite freezing or room temperature storage for 24 h. Our results indicate that ELF collection using Leukosorb paper sampling of ELF provides a sensitive, easy-to-use, and reproducible methodology to collect concentrated amounts of soluble biomarkers from the nasal mucosa. Moreover, the methodology described herein improves upon the standard NL collection method and provides researchers with a novel tool to assess changes in nasal mucosal host defense status.

Influence of cooling face masks on nasal air conditioning and nasal geometry.

INTRODUCTION: Nasal geometries and temperature of the nasal mucosa are the primary factors affecting nasal air conditioning. Data on intranasal air conditioning after provoking the trigeminal nerve with a cold stimulus simulating the effects of an arctic condition is still missing. The objective was to investigate the influence of skin cooling face masks on nasal air conditioning, mucosal temperature and nasal geometry. METHODS: Standardized in vivo measurements of intranasal air temperature, humidity and mucosal temperature were performed in 55 healthy subjects at defined detection sites before and after wearing a cooling face mask. Measurements of skin temperature, rhinomanometry and acoustic rhinometry were accomplished. RESULTS: After wearing the face mask the facial skin temperature was significantly reduced. Intranasal air temperature did not change. Absolute humidity and mucosal temperature increased significantly. The acoustic rhinometric results showed a significant increase of the volumes and the cross-sectional areas. There was no change in nasal airflow. CONCLUSIONS: Nasal mucosal temperature, humidity of inhaled air, and volume of the anterior nose increased after application of a cold face mask. The response is mediated by the trigeminal nerve. Increased mucosal temperatures as well as changes in nasal geometries seem to guarantee sufficient steady intranasal nasal air conditioning.

[Age dynamics of secretory, absorptive, buffer ability of the mucosa inside the human nose].

The problem of age-related functional changes of organs and systems of the human body is one of the urgent problems of modern physiology and gerontology. The most important, complex system of human body is respiratory system, especially its «first line of defense¼ - the nasal cavity. Functional parameters of the nasal cavity undergo a number of specific changesat the stages of postnatal ontogenesis, the study of which has not only scientific value but also practical value for experimental physiology and clinical medicine. The article presents data on the age dynamics and adaptability of such functional parameters of the mucous membrane of the nasal cavity of the person, as secretory and suction activity, the air heater function, the buffer properties by endonasal secret. These indicators are examined at the stages of postnatal development in a comparative perspective based on the study of the age gradients.

Expression of importin-α isoforms in human nasal mucosa: implication for adaptation of avian influenza A viruses to human host.

BACKGROUND: Transportation into the host cell nucleus is crucial for replication and transcription of influenza virus. The classical nuclear import is regulated by specific cellular factor, importin-α. Seven isoforms of importin-α have been identified in human. The preference of importin-α3 of avian influenza virus and -α7 isoform of human strains during replication in human cells was previously identified. In addition, both avian and human influenza viruses were shown to use importin-α1 isoform for their replication. FINDING: The mRNA levels of importin-α1, -α3, and -α7 isoforms in human respiratory tract was determined by real-time RT-PCR. The results indicate that mRNA level of importin-α7 was significantly higher than that of importin-α1 (p-value < 0.0001) and importin-α3 (p-value < 0.0001) isoforms in human nasal mucosa while importin-α1 was detected as the highest expression importin-α isoform in lung tissues. CONCLUSIONS: These results may explain the preference of importin-α7 isoforms in seasonal influenza viruses in human upper respiratory tract and may suggest a selective pressure toward importin-α7 in human respiratory tract infection of an avian virus.

Primary Air-Liquid Interface Culture of Nasal Epithelium for Nasal Drug Delivery.

Nasal drug administration is a promising alternative to oral and parenteral administration for both local and systemic delivery of drugs. The benefits include its noninvasive nature, rapid absorption, and circumvention of first pass metabolism. Hence, the use of an in vitro model using human primary nasal epithelial cells could be key to understanding important functions and parameters of the respiratory epithelium. This model will enable investigators to address important and original research questions using a biologically relevant in vitro platform that mimics the in vivo nasal epithelial physiology. The purpose of this study was to establish, systematically characterize, and validate the use of a primary human nasal epithelium model cultured at the air-liquid interface for the study of inflammatory responses and drug transport and to simultaneously quantify drug effects on ciliary activity.

MgNd2 alloy in contact with nasal mucosa: an in vivo and in vitro approach.

Biodegradable and biocompatible magnesium alloys appear to be very promising not only for temporary clinical application but also for developing deformable and degradable medical implants. This study analyzes the in vivo degradation behavior and the impact on the paranasal sinuses of the highly ductile Mg-2 wt%Nd alloy (MgNd2) in order to provide a basis for a satisfying stent system for the therapy of a chronic sinusitis. Moreover, in vitro tests were carried out on primary porcine nasal epithelial cells (PNEC). For the in vivo tests, cylindrical MgNd2 specimens were implanted into the sinus' mucosa of minipigs. During and after a total period of 180 days the long-term biodegradation and biocompatibility properties after direct contact with the physiological tissue were analyzed. Biodegradation was investigated by measuring the mass and volume losses of the MgNd2 specimens as well as by performing element analyses to obtain information about the degradation layer. The influence on the surrounding tissue of paranasal sinuses was evaluated by endoscopic and histopathological examinations of the mucosa. Here, only a locally unspecific chronic infection was found. The degradation rate showed a maximum after 45 days postsurgery and was determined to decrease subsequently. In vitro experiments using PNEC showed adequate biocompatibility of MgNd2. This study demonstrates a good in vivo biocompatibility for MgNd2 in the system of paranasal sinuses and underlines the promising properties of alloy MgNd2 for biodegradable nasal stent applications.

Mupirocin reduces ciliary beat frequency of human nasal epithelial cells.

Mupirocin is used worldwide for topical treatment of infected skin lesions, impetigo, and especially for nasal decolonization of patients with carriage of Staphylococci, including methicillin-resistant Staphylococcus aureus. Nevertheless, data regarding the effects of mupirocin on the nasal mucosa, in particular on ciliary beat frequency (CBF), is lacking to date. We tested the CBF of ciliated nasal epithelial cells under the influence of Mupirocin-calcium dissolved in tert-butyl alcohol (TBA) containing media in different concentrations comparable to clinical use. Ringer's lactate solution and TBA served as negative control. Cells were visualized with a phase contrast microscope, and the CBF was measured with the SAVA system's region of interest method. Mupirocin-calcium dissolved in TBA led to a statistically significant time- and concentration-dependent decrease in CBF compared to the negative control. TBA addition without mupirocin also led to a significant decrease in CBF, although to a lesser extent than mupirocin/TBA. In conclusion, CBF of human nasal epithelia is significantly reduced by mupirocin-calcium-containing solutions in therapeutic concentrations. Due to our results in this study, mupirocin as a nasal decolonization agent should be used only with care, with a strictly set medical indication, and additional care measures should be considered.

Vanin-1 expression and methylation discriminate pediatric asthma corticosteroid treatment response.

BACKGROUND: There is considerable heterogeneity in asthma treatment response. OBJECTIVE: We sought to identify biomarkers of corticosteroid treatment response in children with asthma and evaluate the utility and mechanistic basis of these biomarkers. METHODS: Children (5-18 years) presenting to the emergency department with an acute asthma exacerbation were recruited and followed during hospitalization. Nasal epithelial cells were collected on presentation to the emergency department (T0) and 18 to 24 hours later (T1), and T1/T0 gene expression ratios were analyzed to identify genes associated with good and poor corticosteroid treatment response phenotypes. The utility of these genes in discriminating between systemic corticosteroid treatment response groups was then tested prospectively in a new cohort of patients. A gene candidate (vanin-1 [VNN1]) that consistently distinguished good versus poor response phenotypes was further studied in an experimental asthma model, and VNN1 promoter methylation was measured by means of bisulfite pyrosequencing in patients. RESULTS: VNN1 mRNA expression changes were associated with systemic corticosteroid treatment response in children with acute asthma, and VNN1 was required for optimal response to corticosteroid treatment in an experimental asthma model. A CpG site within the VNN1 promoter was differentially methylated between good versus poor treatment response groups, and methylation at this site correlated with VNN1 mRNA expression. CONCLUSIONS: We have identified a biological basis for poor corticosteroid treatment response that can be used to distinguish a subgroup of asthmatic children who respond poorly to systemic corticosteroid treatment. VNN1 contributes to corticosteroid responsiveness, and changes in VNN1 nasal epithelial mRNA expression and VNN1 promoter methylation might be clinically useful biomarkers of treatment response in asthmatic children.

Development of glandular models from human nasal progenitor cells.

Hyperplasia/hypertrophy of submucosal glands contributes to mucus overproduction in chronic diseases of the upper and lower respiratory tracts, especially in adult and pediatric chronic rhinosinusitis. Mechanisms that lead to glandular hyperplasia/hypertrophy are markedly understudied, reflecting a lack of in vitro model systems wherein airway epithelial progenitor cells differentiate into glandular cells. In this study, we developed and compared several in vitro three-dimensional systems using human nasal epithelial basal cells (HNEBCs) cultured by different methods on two types of extracellular matrices. We demonstrate that HNEBCs cultured on Matrigel (Corning, Tewksbury, MA) form glandular acini-like structures, whereas HNEBCs embedded in a collagen type I matrix form a network of tubules. Fibroblast-conditioned medium increases tubule formation in collagen type I. In contrast, HNEBCs cocultured with fibroblasts self-aggregate into organotypic structures with tubules and acini. These observations provide morphological evidence that HNEBCs are pluripotent and retain the capacity to differentiate into structures resembling specific structural components of submucosal glands depending on the extracellular matrices and culture conditions. The resultant models should prove useful in targeting cross-talk between epithelial cells and fibroblasts to decipher molecular mechanisms and specific signals responsible for the development of glandular hyperplasia/hypertrophy, which in turn may lead to new therapeutic strategies for chronic rhinosinusitis and other inflammatory respiratory diseases characterized by glandular hyperplasia/hypertrophy.