Increased B Cell-Activating Factor Expression Is Associated with Postoperative Recurrence of Chronic Rhinosinusitis with Nasal Polyps

Heterogeneous inflammatory patterns in chronic rhinosinusitis without nasal polyps in Chicago, Illinois

BackgroundChronic rhinosinusitis (CRS) current therapeutic approaches still fail in some patients with severe persistent symptoms and recurrences after surgery. We aimed to evaluate the master transcription factors gene expression levels of T cell subtypes in chronic rhinosinusitis with nasal polyps (CRSwNP) and chronic rhinosinusitis without nasal polyps (CRSsNP) that could represent new, up-stream targets for topical DNAzyme treatment.Patients and methodsTwenty-two newly diagnosed CRS patients (14 CRSwNP and 8 CRSsNP) were prospectively biopsied and examined histopathologically. Gene expression levels of T-box transcription factor (T-bet, TBX21), GATA binding protein 3 (GATA3), Retinoic acid-related orphan receptor C (RORC) and Forkhead box P3 (FOXP3) were analyzed by real-time quantitative polymerase chain reaction (RT-qPCR).ResultsEosinophilic CRSwNP was characterized by higher level of GATA3 gene expression compared to noneosinophilic CRSwNP, whereas there was no difference in T-bet, RORC and FOXP3 between eosinophilic and noneosinophilic CRSwNP. In CRSsNP, we found simultaneous upregulation of T-bet, GATA3 and RORC gene expression levels in comparison to CRSwNP; meanwhile, there was no difference in FOXP3 gene expression between CRSwNP and CRSsNP.ConclusionsIn eosinophilic CRSwNP, we confirmed the type 2 inflammation by elevated GATA3 gene expression level. In CRSsNP, we unexpectedly found simultaneous upregulation of T-bet and GATA3 that is currently unexplained; however, it might originate from activated CD8+ cells, abundant in nasal mucosa of CRSsNP patients. The elevated RORC in CRSsNP could be part of homeostatic nasal immune response that might be better preserved in CRSsNP patients compared to CRSwNP patients. Further data on transcription factors expression rates in CRS phenotypes are needed.

Chronic rhinosinusitis with nasal polyp (CRSwNP) patients are often characterized by asthma comorbidity and a type-2 inflammation of the sinonasal mucosa. The mucosal microbiota has been suggested to be implicated in the persistence of inflammation, but associations have not been well defined. To compare the bacterial communities of healthy subjects with CRSwNP patients, we collected nasal swabs from 17 healthy subjects, 21 CRSwNP patients without asthma (CRSwNP−A), and 20 CRSwNP patients with co-morbid asthma (CRSwNP+A). We analysed the microbiota using high-throughput sequencing of the bacterial 16S rRNA. Bacterial communities were different between the three groups. Haemophilus influenzae was significantly enriched in CRSwNP patients, Propionibacterium acnes in the healthy group; Staphylococcus aureus was abundant in the CRSwNP−A group, even though present in 57% of patients. Escherichia coli was found in high amounts in CRSwNP+A patients. Nasal tissues of CRSwNP+A patients expressed significantly higher concentrations of IgE, SE-IgE, and IL-5 compared to those of CRSwNP−A patients. Co-cultivation demonstrated that P. acnes growth was inhibited by H. influenzae, E. coli and S. aureus. The nasal microbiota of healthy subjects are different from those of CRSwNP−A and CRSwNP+A patients. However, the most abundant species in healthy status could not inhibit those in CRSwNP disease.

The aim of this study was, firstly, to classify patients with chronic rhinosinusitis with nasal polyps (CRSwNP) and chronic rhinosinusitis without nasal polyps (CRSsNP) into inflammatory endotypes and, secondly, to investigate the relationships between blood and tissue eosinophilia and clinical outcomes in CRSwNP, including disease severity, and recurrence. This cross-sectional, observational, nonrandomized study has been realized between January 2023-December 2024 enrolling a sample of 469 consecutive patients with chronic rhinosinusitis, of which 239 patients with CRSwNP and 230 patients with CRSsNP. The classification of CRSwNP and CRSsNP into inflammatory endotypes was performed by analyzing the levels of blood and tissue eosinophilia. The prevalence of asthma and allergy to nonsteroidal anti-inflammatory drugs (NSAIDs) in the two groups was also analyzed. In the group of patients with CRSwNP, the severity of the disease and the level of quality of life impairment were evaluated, depending on the degree of tissue eosinophilia. The importance of the association between blood vs. tissue eosinophilia and the relapse of CRSwNP was also assessed. The proportion of patients with elevated blood eosinophil levels was higher in CRSwNP than CRSsNP (17.4% vs. 4.35%). Tissue eosinophilia was identified in 163 of 235 (69.4%) patients with CRSwNP compared with 29 of 230 (12.6%) patients with CRSsNP. From an inflammatory perspective, revision surgery was strongly associated with tissue eosinophilia. In contrast, elevated blood eosinophil levels were only moderately associated with revision surgery.

Distinct immunopathologic characteristics of various types of chronic rhinosinusitis in adult Chinese

Conclusion: Biofilms were more prevalent in patients with CRSwNP compared to both CRSsNP and controls, and also on the ethmoid bulla compared to the middle turbinate, supporting a biofilm-related pathogenesis of CRSwNP.Objective: To investigate the prevalence of biofilms in patients with chronic rhinosinusitis with nasal polyps (CRSwNP) compared to patients with chronic rhinosinusitis without nasal polyps (CRSsNP) and controls. To examine the prevalence of biofilms in different anatomical localizations.Study design: Cross-sectional.Methods: This study comprised 27 patients with CRSsNP, 34 patients with CRSwNP, and 25 controls. Biopsies from the middle turbinate, the uncinate process, and the ethmoid bulla were harvested pre-operatively, snap frozen in isopentane, cooled, and stored at −80°C. Prepared with Invitrogens’ Baclight LiveDead kit and investigated with confocal scanning laser microscopy.Results: Biofilms were studied in 33/34 (97%) CRSwNP, 22/27 (82%) CRSsNP, and 14/25 (56%) controls. The difference in point prevalence between patients with CRSwNP vs CRSsNP (p = 0.042, χ2 = 4.12), CRSwNP vs Controls (p < 0.001, χ2 = 15.0), and CRSsNP vs controls (p = 0.047, χ2 = 3.96) were all significant. Biofilms were found in 43/54 (80%) ethmoid bulla, 39/55 (71%) uncinate process, and 31/50 (62%) middle turbinate. The difference between the ethmoid bulla and the middle turbinate locations (p = 0.047, χ2 = 3.93) was significant.

Serine proteinase inhibitors, clade B, member 3 (SerpinB3) and B4 are highly similar in amino acid sequences and associated with inflammation regulation. We investigated SerpinB3 and B4 expression and their roles in chronic rhinosinusitis with nasal polyps (CRSwNP). The expression of SerpinB3 and B4 in nasal mucosa tissues, brush cells, and secretions from CRSwNP patients was measured, and their regulation by inflammatory cytokines were investigated. Their functions were also analyzed using air-liquid interface (ALI)-cultured primary human nasal epithelial cells (HNECs) and transcriptomic analysis. Both SerpinB3 and B4 expression was higher in nasal mucosa, brush cells, and secretions from eosinophilic (E) CRSwNP and nonECRSwNP patients than in healthy controls. Immunofluorescence staining indicated that SerpinB3 and B4 were primarily expressed in epithelial cells and their expression was higher in CRSwNP patients. SerpinB3 and B4 expression was upregulated by interleukin-4 (IL-4), IL-5, IL-6, and IL-17a. Transcriptomic analysis identified differentially expressed genes (DEGs) in response to recombinant SerpinB3 and B4 stimulation. Both the DEGs of SerpinB3 and B4 were associated with disease genes of nasal polyps and inflammation in DisGeNET database. Pathway enrichment indicated that downregulated DEGs of SerpinB3 and B4 were both enriched in cytokine-cytokine receptor interactions, with CXCL8 as the hub gene in the protein-protein interaction networks. Furthermore, CXCL8/IL-8 expression was downregulated by recombinant SerpinB3 and B4 protein in ALI-cultured HNECs, and upregulated when knockdown of SerpinB3/B4. SerpinB3/B4 expression is upregulated in nasal mucosa of CRSwNP patients. SerpinB3/B4 may play an anti-inflammatory role in CRSwNP by inhibiting the expression of epithelial cell-derived CXCL8/IL-8.

Objective:To study the expression and role of interleukin-18 (IL-18) in different clinical phenotypes of chronic rhinosinusitis(CRS)and in different subtypes of CRS with nasal polyps (CRSwNP).Method:During nasal endoscopic surgery, inferior turbinates were obtained from 13 patients with nasal septum deviation (control group),uncinate processes were obtained from 10 patients with chronic rhinosinusitis without nasal polyps (CRSsNP) and nasal polyp tissues were obtained from 36 patients with CRSwNP, respectively. IL-5 expression in CRSwNP was detected by ELISA, and the expression of IL-18 mRNA and protein in different subtypes of CRS were assessed by real-time PCR, ELISA and immunohistochemistr.Result:CRSwNP was divided into 12 cases of IL-5 positive nasal polyps (IL-5+NP)group and 24 cases of IL-5 negative nasal polyps (IL-5-NP)group. The expression of IL-18 mRNA and protein in all CRSsNP,IL-5+NP and IL-5-NP groups were higher than that in control group (P<0.01 or P<0.05),but no significant difference existed between IL-5+NP and IL-5-NP (P>0.05) group. IL-18 also expresses in epithelial cells of normal nasal mucosa.Conclusion:CRSwNP can be divided into two main subtypes: IL-5+NP and IL-5-NP. The increasing expression of IL-18 in CRSsNP, IL-5+NP and IL-5-NP compared with control group indicates that IL-18 may play a key role in the pathogenesis of CRS, and IL-18 expression in nasal polyps is not affected by the type of inflammation. The possible presence of balance between IL-18 and IL-18 binding protein in normal nasal mucosa provides a new way for research and treatment of CRS.

Chronic rhinosinusitis with nasal polyps (CRSwNP) is a chronic sinonasal disease characterized by heterogeneous inflammation. However, the presence of systemic inflammation heterogeneity in CRSwNP patients remains unknown. This study aims to profile transcriptomic alterations in the blood of CRSwNP patients and characterize the CRSwNP heterogeneity based on blood transcriptomic biomarkers. Patients with CRSwNP were prospectively recruited from three hospitals and chronologically divided into exploratory (n=123) and independent validation (n=46) cohorts. Transcriptomic profiles were generated by whole blood mRNA sequencing and subjected to patient clustering, differential expression, and pathway analysis. Differences in immune pattern and clinicopathologic features between clusters were assessed. A transcriptomic signature was defined and applied to an independent cohort to validate the findings. CRSwNP patients showed diverse blood transcriptomic profiles versus healthy controls, or when stratified by tissue and blood eosinophils and asthma comorbidity. Transcriptome-wide correlation analysis revealed a transcriptional signature associated with blood eosinophil levels, consisting of nine T2-related genes (CLC, SIGLEC8, ALOX15, IL5RA, PTGDR2, CCL23, CCR3, EPX and IL1RL1). Three distinct clusters with differing systemic eosinophilic and neutrophilic inflammation patterns and asthma comorbidity were identified based on transcriptomic profiling of T2 and T1/3-related blood biomarkers. A 36-gene signature was developed by machine learning and accurately predicted the three CRSwNP subtypes. Validation on an independent cohort confirmed the prediction robustness. There is heterogeneous systemic inflammation associated with eosinophilic and neutrophilic patterns in patients with CRSwNP. Endotyping based on blood transcriptomic biomarkers might lead to more personalized treatment strategies for CRSwNP in the future.

Background: As of today, chronic rhinosinusitis (CRS) characterized by inflammatory processes and tissue remodeling of the mucosal lining is classified into two phenotypes: chronic rhinosinusitis with nasal polyps (CRSwNP) and chronic rhinosinusitis without nasal polyps (CRSsNP). Matrix metalloproteinases (MMPs) constitute a family of zinc-dependent endopeptidases that play a crucial role in degrading basement membranes and extracellular matrix. MMP-7 and MMP-9 may play significant roles in CRS development. Objective: To determine the level of zinc-dependent endopeptidases in the tissues of patients with CRSwNP and CRSsNP. Materials and Methods: The study included 54 patients meeting the inclusion criteria, among them 12 patients in the control group, and 21 patients each in subgroup A (CRSwNP) and subgroup B (CRSsNP) of the main group. Patients underwent SNOT-22 questionnaire, endoscopic evaluation using the Lund-Kennedy scale, CT scans graded by the Lund-Mackay scale, and levels of MMP-9 and MMP-7 enzymes were determined in surgically removed fragments of pathological tissues. Results: It was found that CRSwNP is more commonly associated with olfactory dysfunction. However, obstruction of the olfactory cleft by polypoid elements, according to CT scans, was observed in only 19.04% of cases (4 out of 21 patients). The level of MMP-7 for the control group was 8.17±5.15 μg/ml, for subgroup A it was 23.67±11.89 μg/ml, and for subgroup B it was 17.93±9.88 μg/ml. Student's t-test revealed no significant difference between subgroups A and B of the main group, p&gt;0.05 (T-statistic: 1.7; p-value: 0.09). However, when compared individually with the control group, a statistical difference was observed in each case, p&lt;0.05. In contrast to MMP-7 data, statistically significant differences were observed when comparing the MMP-9 levels of each group using the t-test, p&lt;0.05. The distribution of mean MMP-9 levels for the groups was as follows: control group – 17.49±8.81 μg/ml, subgroup A – 36.04±16.03 μg/ml, and subgroup B – 26.06±11.04 μg/ml. Conclusions: MMP-9 and MMP-7 are key enzymes involved in the formation of polypoid tissue or pseudocysts in different phenotypes of CRS and influence the severity of the disease's clinical course. Therefore, these endopeptidases could potentially serve as biomarkers for CRS activity and as targets for new therapeutic agents that could inhibit them. Keywords: chronic rhinosinusitis, endoproteases, MMP-9, MMP-7, nasal cavity, nasal mucosa, nasal polyps, nasal cavity pseudocysts.

Tezepelumab treatment in severe asthma with recurrent chronic rhinosinusitis with nasal polyps: Case series.

BackgroundThe conventional belief that glucocorticosteroid (GC) acts through a single brand glucocorticoid receptor (GR)α protein has changed dramatically with the discovery of multiple GR isoforms. We aimed to evaluate whether multiple GR protein isoforms are expressed in chronic rhinosinusitis with nasal polyps (CRSwNP) and whether GR protein isoform expression profiles differ between different endotypes of CRSwNP.MethodsThirty-eight patients with CRSwNP and ten healthy volunteers were included. The protein expression of multiple GR isoforms in nasal polyps (NPs) tissue and control mucosae was examined by western blot analysis with different GR antibodies.ResultsFive bands, including three bands for known proteins (GRα-A/B, GRα-C, and GRα-D) and two bands for unidentified proteins at 67 kilodaltons (kDa) and 60 kDa, were identified with both total GR antibody (PA1-511A) and GRα-specific antibody (PA1-516). GRα-D intensity, which was abundant in nasal mucosa, was significantly increased in the CRSwNP group and was especially elevated in the noneosinophilic CRSwNP (NE-CRSwNP) group (PA1-511A: P < 0.001 and P = 0.0018; PA1-516: P < 0.003 and P = 0.006, respectively). Additionally, the intensities of the newly recognized 67 kDa and 60 kDa bands were much greater in the NE-CRSwNP subgroup than in the eosinophilic CRSwNP (E-CRSwNP) subgroup; in the E-CRSwNP subgroup, the median intensities were even lower than those in the control group.ConclusionsThis study provides evidence that nasal tissues express multiple GR protein isoforms. GR protein isoforms presented disease and tissue-specific expression profiles that differed between the CRSwNP and control groups and between the E-CRSwNP and NE-CRSwNP subgroups.Graphical abstract

microRNAs (miRNAs) are directly associated with inflammatory response, but their direct role in CRSwNP (chronic rhinosinusitis with nasal polyps) remains evasive. This study aimed to compare the expression of several miRNAs in tissue samples obtained from patients with CRSwNP and controls and to evaluate if miRNAs correlate to a specific inflammatory pattern (T1, T2, T17, and Treg) or intensity of symptoms in CRSwNP. nasal polyps (from patients with CRSwNP - n=36) and middle turbinate mucosa (from control patients - n=41) were collected. Microarray determined human mature miRNA expression, and the results obtained were validated by qPCR. miRNAs that were differentially expressed were then correlated to cytokine proteins (by Luminex), tissue eosinophilia, and SNOT-22. After microarray and qPCR analyses, six microRNAs were up-regulated in CRSwNP samples when compared with controls: miR-205-5p, miR-221-3p, miR-222-3p, miR-378a-3p, miR-449a and miR-449b-5p. All these miRNAs are directly implicated with cell cycle regulation and apoptosis, and to a minor extent, with inflammation. Importantly, miR-205-5p showed a significantly positive correlation with IL-5 concentration and eosinophil count at the tissue and with the worst SNOT-22 score. miRNA 205-5p was increased in CRSwNP compared to controls, and it was especially expressed in CRSwNP patients with higher T2 inflammation (measured by both IL-5 levels and local eosinophilia) and worst clinical presentation. This miRNA may be an interesting target to be explored in patients with CRSwNP.

The antimicrobial protein short palate, lung, and nasal epithelium clone 1 (SPLUNC1) is differentially modulated in eosinophilic and noneosinophilic chronic rhinosinusitis with nasal polyps

To investigate the clinical significance of specific IgE-staphylococcal enterotoxin B (IgE-SEB) in CRS (chronic rhinosinusitis). Retrospective analysis of patients who were positive for specific IgE-staphylococcal enterotoxin B. Tertiary rhinology clinic. A total of 965 patients who were tested for specific IgE-staphylococcal enterotoxin B from December 2016 to December 2017. We retrospectively reviewed the records of 965 patients who were tested for specific IgE-staphylococcal enterotoxin B from December 2016 to December 2017. Patient demographics, titre-specific IgE to staphylococcal enterotoxin B levels, MAST, serologic test and medical records were reviewed. IgE-SEB (KU/L) was higher in CRS patients than non-CRS patients (0.13±0.37 vs 0.08±0.22, respectively; P-value: .044), and the IgE-SEB (+, ≥0.35) rate was also higher (10.06% vs 4.46%, respectively; P-value: .030). IgE-SEB(KU/L) was higher in the CRS group than in the fungal sinusitis group (0.13±0.37 vs 0.03±0.05, respectively; P-value: <.001), and the IgE-SEB (+, ≥0.35) rate was also higher (10.06% vs 0%, respectively; P-value: .015). Between the CRSsNP (chronic rhinosinusitis without nasal polyps) and CRSwNP (chronic rhinosinusitis with nasal polyps) groups, there were no differences in IgE-SEB (KU/L) or IgE-SEB (+) rates. IgE-SEB positivity was not associated with the presence of polyps, concomitant asthma or postoperative recurrence. As the values of IgE-SEB (KU/L) and the IgE-SEB (+, >0.1) rate increased, the CRS severity also increased. IgE-SEB showed a positive correlation with Lund-Mackay CT severity score, but not with postoperative recurrence or nasal polyps. Further studies are needed to obtain clear evidence that IgE-SEB can be considered as an independent CRS endotype.