Pathogenic role of different phenotypes of immune cells in airway allergic diseases: a study based on Mendelian randomization.

Background: Airway allergic disease (AAD) is a class of autoimmune diseases with predominantly Th2-type inflammation, mainly including allergic rhinitis (AR), allergic asthma (AS), and chronic sinusitis (CRS). There are very complex regulatory mechanisms between immune cells and AAD; however, previous reports found that the functions of the same immune cells in AAD are not identical. Objective: The aim of this study was to explore the causal relationship between different phenotypic immune cells and their association with AAD. Method: Utilizing the publicly available Genome-Wide Association Studies (GWAS) database, this study conducted a bidirectional Mendelian randomization (MR) to assess the causal relationship between immune cells of 731 different immunophenotypes and AAD. The primary assessment methods included inverse variance weighting, weighted median, and MR Egger. Additionally, sensitivity analyses such as MR-PRESSO, leave-one-out, and scatter plots were employed to eliminate the interference of heterogeneity and pleiotropy, ensuring the stability of the causal inference. Result: A total of 38 immune cells with different immunophenotypes were found to be positively and causally associated with AR, of which 26 were protective factors and 12 were risk factors. Positive associations were found between 33 immune cells and AS, of which 14 were protective factors and 19 were risk factors, as well as between 39 immune cells and CRS, of which 22 were protective factors and 17 were risk factors. Finally, the results of all relevant immune cells for the three diseases were taken and intersected, and it was found that CD3 on CD39+-activated Treg (IVWAR = 0.001, IVWCRS = 0.043, IVWAS = 0.027) may be the key immune cell that inhibits the development of AAD (ORAR = 0.940, ORAS = 0.967, ORCRS = 0.976). Conclusion: This study reveals that different immune phenotypes of immune cells are closely related to AAD at the genetic level, which provides a theoretical basis for future clinical studies.

The role of ZEB1 in regulating tight junctions in antrochoanal polyp.

Background: Antrochoanal polyp (ACP) is a benign nasal mass of unknown etiology. Tight junctions (TJs) are essential to the epithelial barrier that protects the body from external damage. However, the phenotype of tight junction in ACP is currently unclear. Methods: The samples were collected from 20 controls, 37 patients with ACP and 45 patients with chronic rhinosinusitis with nasal polyp (CRSwNP). Quantitative Real-Time PCR (qRT-PCR) and immunofluorescence staining (IF) were performed to analyze the expressions of TJs markers (ZO-1, claudin-3 and occludin) and ZEB1. hNEpCs were transfected with ZEB1 small interfering RNA (si-ZEB1) or ZEB1 over-expression plasmid (OE-ZEB1). qRT-PCR and Western blotting were used to determine the levels of TJs-related markers. Primary human nasal epithelial cells (hNECs) were stimulated with IL-17A and si-ZEB1, and the expression of epithelial barrier markers were measured by qRT-PCR and Western blotting. Results: Compared to the control group, ACP group showed a significant downregulation in both mRNA and protein levels of ZO-1, occludin, and claudin-3. Furthermore, disease severity correlates positively with the degree of disruption of tight junctions. In addition, higher expression levels of ZEB1, IL-17A, and IFN-γ were observed in the ACP group compared to controls. Overexpression of ZEB1 in hNEpCs led to impairments in the levels of ZO-1, occludin, and claudin-3, while silencing of ZEB1 expression was found to enhance the barrier function of epithelial cells. Finally, IL-17 stimulation of hNECs impaired the expression of TJs-associated molecules (ZO-1, occludin, and claudin-3), which was effectively reversed by the IL-17A + si-ZEB1 group. Conclusions: The tight junctions in ACP were extremely damaged and were correlated with the severity of the disease. ZEB1 was involved in the pathogenesis of ACP mediated by IL-17A through regulating tight junctions.

miR-200a-3p regulates epithelial-mesenchymal transition and inflammation in chronic rhinosinusitis with nasal polyps by targeting ZEB1 via ERK/p38 pathway.

BACKGROUND: Several biological processes are regulated by miR-200a-3p, including cell proliferation, migration, and epithelial-mesenchymal transition (EMT). In this study we aimed to uncover the diagnostic value and molecular mechanisms of miR-200a-3p in chronic rhinosinusitis with nasal polyps (CRSwNP). METHODS: The expressions of miR-200a-3p were detected by quantitative real-time polymerase chain reaction (qRT-PCR), Zinc finger E-box binding homeobox 1 (ZEB1) levels were examined by qRT-PCR and immunofluorescence staining. The interaction between miR-200a-3p and ZEB1 was predicted by TargetScan Human 8.0 and confirmed by dual-luciferase reporter assays. In addition, the effect of miR-200a-3p and ZEB1 on EMT-related makers and inflammation cytokines was assessed by qRT-PCR and Western blotting in human nasal epithelial cells (hNEpCs) and primary human nasal mucosal epithelial cells (hNECs). RESULTS: We found that miR-200a-3p was downregulated in non-eosinophilic and eosinophilic CRSwNP patients when compared with controls. The diagnostic value of miR-200a-3p in serum is reflected by the receiver operating characteristic curve and the 22-item Sino-Nasal Outcome Test. Bioinformatic analysis and luciferase reporter assay identified ZEB1 as a target of miR-200a-3p. ZEB1 was more highly expressed in CRSwNP than in controls. Furthermore, miR-200a-3p inhibitor or ZEB1 overexpression significantly suppressed the epithelial marker E-cadherin; promoted the activation of vimentin, α-spinal muscle atrophy, and N-cadherin; and aggravated inflammation in hNEpCs. Knockdown of ZEB1 significantly alleviated the cellular remodeling caused by miR-200a-3p inhibitor via the extracellular signal-regulated kinase (ERK)/p38 pathway in hNECs. CONCLUSIONS: miR-200a-3p suppresses EMT and inflammation by regulating the expression of ZEB1 via the ERK/p38 pathway. Our study presents new ideas for protecting nasal epithelial cells from tissue remodeling and finding a possible target for disease.

Difference of Antrochoanal Polyps Between Children and Adults in the Chinese Population.

OBJECTIVE: This study aims to find the difference in clinical and immunopathological characteristics between children and adults with antrochoanal polyps (ACPs) in the Chinese population. METHODS: The clinical data of 69 patients diagnosed with ACPs were retrospectively analyzed. Cytokine levels in 16 controls and 40 ACPs tissues were determined by quantitative real-time polymerase chain reaction (qPCR). The expression of matrix metalloproteinase (MMP)-9 was measured using qPCR, immunofluorescent staining, and western blot. RESULTS: There were 51 (73.9%) children (<18 years old) and 18 (26.1%) adults (≥18 years old). The sex ratio differed significantly between the two groups (p = 0.0032). There were no significant differences in the nasal side of ACPs and approaches to surgery between the two groups. In both groups, the most common symptom was nasal obstruction, followed by nasal discharge. As for associated nasal diseases, there was a significant difference between the two groups in septal deviation (p = 0.0223). Adult patients showed significantly higher expression of IL-8 mRNA than children (p = 0.0424). The mRNA and protein levels of MMP-9 were also significantly higher in adult patients than in children (p = 0.0498 and 0.0009, respectively). CONCLUSION: In the Chinese population, the comorbidities and immunopathological characteristics of adult ACP patients are different from those of children. The level of IL-8 and MMP-9 was significantly higher in ACPs of adults than in children, which may contribute to the more severe tissue remolding in adult ACP patients. LEVEL OF EVIDENCE: 3 Laryngoscope, 134:2093-2099, 2024.

Predictive Value of Adenoid-Nasopharyngeal Ratio in the Diagnosis of Secretory Otitis Media.

OBJECTIVES: To explore the associated factors of otitis media with effusion (OME) and analyze the diagnostic value of the adenoid-nasopharyngeal (A/N) ratio to OME. METHODS: Patients with adenoid hypertrophy (AH) recently in 2 years were collected, including sex, age, duration, with/without rhinosinusitis, and examination results, including Cone Beam Computerized Tomography (CBCT) sinus imaging, tympanometry, pure tone audiometry (PTA), blood test, and allergen detection. According to Liden/Jerger's classification, the patients were divided into two groups: Type B and type C, which were defined as the tympanometry abnormal group (TAG), and the rest were classified as the tympanometry normal group (TNG). RESULTS: A total of 316 children were included in this study. Age and duration were significantly younger and shorter in TAG (6.0(4.0-9.0) vs.5.0(4.0-7.0)); 12.0(4.0-24.0) vs.6.0(2.0-12.0)). Compared to TNG, the allergen test results of fx5 (protein, milk, cod, wheat, peanut, and soybean) in TAG were higher (0.09(0.04-0.25) vs.0.14(0.05-0.45)), but eosinophilia in blood was lower (count: 0.21(0.13-0.35) vs. 0.18(0.12-0.27); ratio: 3.10 (1.90-4.70) vs. 2.50 (1.65-3.80)). A/N ratio and Visual obstruction ratio had a statistical difference (Z = -3.770, P < .01) but the two ratios didn't have too much disparity (0.82(0.74-0.88) VS 0.80(0.75-0.80)), and they had a positive correlation (r = 0.345, P < .01). A/N ratio of TAG was higher than TNG (0.78(0.70-0.85) vs. 0.86(0.82-0.90)) and had a positive correlation with increasing negative middle ear pressure (r = -3.777, P < .01). A/N ratio was an associated factor of OME (OR:1355.611, P = .006), the cut-off value of A/N ratio was 0.815(sensitivity: 75.3%, specificity: 64.3%, area under the curve (AUC): 0.747). CONCLUSIONS: A/N ratio indirectly reflected the abnormality of tympanometry in this study. When A/N ratio reaches 0.815, patients are at a higher risk of having OME so it could be a predictor of OME in patients with adenoid hypertrophy.

An Integrated Analysis of Inflammatory Endotypes and Clinical Characteristics in Chronic Rhinosinusitis with Nasal Polyps.

Objective: Chronic rhinosinusitis with nasal polyps (CRSwNP) is mainly characterised by type 1 (T1), type 2 (T2) and type 3 (T3) inflammatory endotypes. However, correlations between inflammatory endotypes and clinical features in CRSwNP have not been demonstrated sufficiently. This study aimed to determine the endotype-phenotype associations in CRSwNP. Methods: Clinical data of 31 control subjects and 106 CRSwNP patients were analysed. Interferon (IFN)-γ (T1), Charcot-Leyden crystal galectin (CLC) (T2) and Interleukin (IL)-17A (T3) were used as biomarkers to identify the inflammatory endotypes. Results: The mRNA expression level of IFN-γ was positively correlated with IL-17A (r = 0.817; P < 0.0001). Headache/facial pain (P = 0.039) was associated with T1 endotype. Smell loss (P = 0.025) was associated with T2 endotype. Purulent rhinorrhea (P = 0.001) was associated with T3 endotype. Atopy (P = 0.030), asthma (P = 0.005) and recurrence (P = 0.022) were more frequent in T2 endotype. Total Symptom Scores (TSS) of T2 (P < 0.001) and T3 (P = 0.009) endotype were higher than non-T2 and non-T3, respectively. Sino Nasal Outcome Test-22 (SNOT-22) total scores of T3 (P = 0.054) endotype were higher than non-T3. Conclusion: Identifications of endotype-phenotype associations are useful in clinical diagnoses and targeted therapies for patients with CRSwNP.