Reduced Expression of Antimicrobial Protein Secretory Leukoprotease Inhibitor and Clusterin in Chronic Rhinosinusitis with Nasal Polyps

Introduction. Antimicrobial peptides and proteins (AMPs) constitute the first line of defense against pathogenic microorganisms in the airway. The association between AMPs and chronic rhinosinusitis with nasal polyps (CRSwNP) requires further investigations. This study is aimed at investigating the expression and regulation of major dysregulated AMPs in the nasal mucosa of CRSwNP. Methods. The expression of AMPs was analyzed in nasal tissue from patients with eosinophilic (E) CRSwNP and nonECRSwNP and healthy subjects using RNA sequencing. The 10 most abundant AMPs expressed differentially in CRSwNP patients were verified by real-time PCR, and of these, the expression and regulation of secretory leukoprotease inhibitor (SLPI) and clusterin (CLU) were investigated further. Results. The 10 most abundant AMPs expressed differentially in CRSwNP compared to healthy control, regardless of subtypes, included BPIFA1, BPIFB1, BPIFB2, CLU, LTF, LYZ, and SLPI, which were downregulated, and S100A8, S100A9, and HIST1H2BC, which were upregulated. ELISA and immunofluorescence confirmed the decreased expression of SLPI and CLU levels in CRSwNP. SLPI is expressed in both nasal epithelial cells and glandular cells, whereas CLU is mainly expressed in glandular cells. AB/PAS staining further demonstrated that both SLPI and CLU were mainly produced by mucous cells in submucosal glands. Furthermore, the numbers of submucosal glands were significantly decreased in nasal polyp tissue of CRSwNP compared to nasal tissue of controls. SLPI was downregulated by TGF-β1 and IL-4 in cultured nasal tissues in vitro, while CLU expression was inhibited by TGF-β1. Glucocorticoid treatment for 2 weeks significantly increased the expression of all downregulated AMPs, but not LYZ. Additionally, budesonide significantly increased the expression of SLPI and CLU in cultured nasal tissues. Conclusion. The expression of major antimicrobial proteins is significantly decreased in nasal tissue of CRSwNP. The expression of SLPI and CLU is correlated with the numbers of submucosal glands and regulated by inflammatory cytokines and glucocorticoids.

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Elevated S100A9 expression in chronic rhinosinusitis coincides with elevated MMP production and proliferation in vitro

Scientific Reports ◽

10.1038/s41598-020-73480-8 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Marina Boruk ◽

Christopher Railwah ◽

Alnardo Lora ◽

Sridesh Nath ◽

Derek Wu ◽

...

Keyword(s):

Gene Expression ◽

Cell Proliferation ◽

Chronic Rhinosinusitis ◽

Nasal Polyps ◽

Plasma Concentrations ◽

Nasal Tissue ◽

Elevated Expression ◽

Proliferation In Vitro ◽

S100a9 Expression

Abstract Chronic rhinosinusitis (CRS) is a common condition associated with inflammation and tissue remodeling of the nose and paranasal sinuses, frequently occurring with nasal polyps and allergies. Here we investigate inflammation and the protease profile in nasal tissues and plasma from control non-CRS patients and CRS patients. Gene expression for several cytokines, proteases, and antiproteases was quantified in nasal tissue from non-CRS and CRS subjects with nasal polyps. Elevated expression of S100A9, IL1A, MMP3, MMP7, MMP11, MMP25, MMP28, and CTSK was observed in tissue from CRS subjects with nasal polyps compared to control tissue. Tissue protein analysis confirmed elevated levels of these targets compared to controls, and increased MMP3 and MMP7 observed in CRS subjects with nasal polyps compared to CRS subjects without polyps. Plasma concentrations of MMP3 and MMP7 were elevated in the CRS groups compared to controls. The nasal cell line, CCL-30, was exposed to S100A9 protein, resulting in increased MMP3, MMP7, and CTSK gene expression and elevated proliferation. Silencing MMP3 significantly reduced S100A9-mediated cell proliferation. Therefore, the elevated expression of S100A9 and MMPs are observed in CRS nasal tissue and S100A9 stimulated MMP3 responses to contribute to elevated nasal cell proliferation.

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CD133, a Progenitor Cell Marker, is Reduced in Nasal Polyposis and Showed Significant Correlations with TGF-β1 and IL-8

International Archives of Otorhinolaryngology ◽

10.1055/s-0041-1726043 ◽

2021 ◽

Author(s):

Wagner Vargas Souza Lino ◽

André Luis Lacerda Bachi ◽

José Arruda Mendes Neto ◽

Gabriel Caetani ◽

Jônatas Bussador do Amaral ◽

...

Keyword(s):

Chronic Rhinosinusitis ◽

Nasal Polyps ◽

Nasal Polyposis ◽

Transforming Growth Factor ◽

Middle Turbinate ◽

Enzyme Linked Immunosorbent Assay ◽

Control Group ◽

Histologic Analysis ◽

Aspirin Intolerance ◽

Tgf Β1

Abstract Introduction Combination of chronic inflammation and an altered tissue remodeling process are involved in the development of Chronic Rhinosinusitis with Nasal Polyps (CRSwNP). Studies demonstrated that mesenchymal stem cells expressing the progenitor gene CD133 were involved in a significant reduction of the chronic inflammatory process in the polypoid tissue. Objective To evaluate the levels of CD133 (Prominin-1) in nasal polypoid tissue and its correlation with interleukin-8 (IL-8) and transforming growth factor β1 (TGF-β1). Methods A total of 74 subjects were divided in the following groups: control group (n = 35); chronic rhinosinusitis with nasal polyps nonpresenting comorbid asthma and aspirin intolerance (CRSwNPnonAI) group (n = 27); and chronic rhinosinusitis with nasal polyps presenting comorbid asthma and aspirin intolerance (CRSwNPAI) group (n = 12). Histologic analysis and also evaluation of the concentration of CD133, IL-8, and TGF-β1 by enzyme-linked immunosorbent assay (ELISA) kits were performed in nasal tissue obtained from nasal polypectomy or from middle turbinate tissue. Results Higher eosinophilic infiltration was found in both CRSwNP groups by histologic analysis. Lower levels of TGF-β1 and IL-8 were observed in both CRSwNP groups when compared with the control group, whereas the CD133 levels were significantly reduced only in the CRSwNPnonAI group compared with the control group. Conclusion It was demonstrated that the nasal mucosa presenting polyposis showed a significant reduction of CD133 levels, and also that this reduction was significantly correlated with the reduction of TGF-β1 levels, but not with IL-8 levels. Therefore, these findings may be involved in the altered inflammatory and remodeling processes observed in the nasal polyposis.

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Increased Expression of TXNIP Facilitates Oxidative Stress in Nasal Epithelial Cells of Patients With Chronic Rhinosinusitis With Nasal Polyps

American Journal of Rhinology and Allergy ◽

10.1177/1945892420982411 ◽

2020 ◽

pp. 194589242098241

Author(s):

Hai Lin ◽

Guangyi Ba ◽

Ru Tang ◽

Mingxian Li ◽

Zhipeng Li ◽

...

Keyword(s):

Oxidative Stress ◽

Epithelial Cells ◽

Real Time ◽

Chronic Rhinosinusitis ◽

Western Blotting ◽

Real Time Pcr ◽

Nasal Polyps ◽

Sod Activity ◽

Nasal Tissue ◽

Sod Assay

Background Oxidative stress plays crucial roles in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP). Thioredoxin-interacting protein (TXNIP) is essential in the process of triggering oxidative stress. However, its role and mechanism in CRSwNP remain unclear. The present study sought to explore the role and mechanism of TXNIP in the pathogenesis of CRSwNP. Methods Western blotting, real-time PCR and immunohistochemistry (IHC) were employed to assess TXNIP, thioredoxin (TRX) expression in nasal tissue samples from patients with CRSwNP and control subjects. MDA level and SOD activity in nasal tissue homogenates were measured using MDA and SOD Assay Kit. To evaluate the role and mechanism of TXNIP in CRSwNP, human nasal epithelial cells (HNECs) were cultured and stimulated using TXNIP siRNA, with or without N-acetylcysteine (NAC, an ROS scavenger). Western blotting, real-time PCR, ROS detecting dye DCFH-DA, MDA and SOD Assay Kit were performed to assess the effects and mechanisms of stimulators on the cells. Results We found significantly increased levels of TXNIP and decreased levels of TRX protein, mRNA, positive cells, increased MDA level and decreased SOD activity in CRSwNP patients compared with control subjects. In vitro study, significantly altered levels of TXNIP, TRX, MDA, SOD and ROS in HNECs were found following treatment of TXNIP siRNA with or without NAC on HNECs. Conclusion TXNIP expression was increased and TRX expression was decreased in CRSwNP at both protein and mRNA levels. MDA levels were increased and SOD activities were decreased in CRSwNP. TXNIP may have negative association with TRX, and then decrease SOD activities and increase MDA levels, resulting in the upregulation of ROS and oxidative stress in HNECs, which may play a pivotal role in the pathogenesis of CRSwNP. Future studies are expected to further explore the role and mechanism of TXNIP in CRSwNP.

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In Vitro and in Vivo Antigen-Induced Release of High-Molecular Weight Neutrophil Chemotactic Activity from Human Nasal Tissue

American Journal of Rhinology ◽

10.2500/105065888781693221 ◽

1988 ◽

Vol 2 (1) ◽

pp. 7-11 ◽

Cited By ~ 4

Author(s):

T. Nagakura ◽

T. Onda ◽

Y. likura ◽

T. Endo ◽

H. Nagakura ◽

...

Keyword(s):

Molecular Weight ◽

High Molecular Weight ◽

Nasal Polyps ◽

Gel Filtration ◽

Chemotactic Activity ◽

Nasal Tissue ◽

Nasal Secretions ◽

Neutrophil Chemotactic Activity

High molecular weight neutrophil chemotactic activity has been identified in resected human nasal polyps, inferior turbinates, and nasal secretions following antigen challenge. The estimated molecular weight, by gel filtration chromatography, was approximately 600,000. However, a heterogeneity of molecular weight in some patients was recognized. Our results suggest a possible role for high molecular weight-neutrophil chemotactic activity in the pathogenesis of hypersensitivity in the human nasal cavity.

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Differences in Expression of Inflammatory Mediator in Mucosal and Polyp Tissue between Chronic Rhinosinusitis and Recurrent Chronic Rhinosinusitis

Open Access Macedonian Journal of Medical Sciences ◽

10.3889/oamjms.2019.341 ◽

2019 ◽

Vol 7 (11) ◽

pp. 1733-1738 ◽

Cited By ~ 1

Author(s):

Effy Huriyati ◽

Eryati Darwin ◽

Yanwirasti Yanwirasti ◽

Irza Wahid

Keyword(s):

Chronic Rhinosinusitis ◽

Nasal Polyps ◽

Surgical Removal ◽

Main Role ◽

Cross Sectional ◽

Significant Difference ◽

Polyp Tissue ◽

The Difference ◽

Tissue Specimens ◽

Tgf Β1

BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) remains a challenging clinical entity with its propensity for recurrence. This disease decreases the patients’ quality of life and creates a high economic burden. An effort to investigate the aetiology of recurrent polyps have to be more alert. AIM: This study aims to prove the differences in expression of IL-5, IL-8, IL-17A and TGF-β1 in mucosal and polyp tissue between CRSwNP and recurrent CRSwNP and also to determine which expression of cytokines that have the main role in mucosal and polyp tissue in recurrent CRSwNP. MATERIAL AND METHODS: An observational study was conducted with a comparative cross-sectional design of CRS patients with 15 recurrent CRSwNP and CRSwNP who had never undergone surgery for as many as 15 polyps. Mucosal specimens of nasal polyps are taken by brushing, and polyp tissue specimens are taken during surgical removal of nasal polyps. Specimens from the polyp mucosa were examined by ELISA while the polyp tissue specimens were carried out immunohistochemistry (IHC). RESULTS: The result showed that there is a significant difference in IL-5 expression in the polyp mucosal between CRSwNP with recurrent CRSwNP, where expression is higher in recurrent CRSwNP. The expression of IL-8, IL-17 and TGF-β1 were lower in recurrent CRSwNP, but the difference was not significant. In nasal polyp tissue, there is a significant difference in TGF-β1 and IL-8 expression between CRSwNP and recurrent CRSwNP, where the expression of both cytokines is lower in recurrent CRSwNP. Interleukin-5 expression was higher in recurrent CRSwNP than CRSwNP, but the difference was not significant. In the polyps mucosal, IL-5 has the main role in recurrent CRSwNP polyp, whereas TGF-β has the main role in polyp tissue. CONCLUSION: This study concluded that the expression of IL-5 in the mucosa could be examined with simple techniques like brushing before polypectomy or FESS was performed to determine the possibility of polyps recurrencies.

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IL-19 Induced by IL-13/IL-17A in the Nasal Epithelium of Patients with Chronic Rhinosinusitis Upregulates MMP-9 Expression via ERK/NF-kB Signaling Pathway

10.21203/rs.3.rs-46150/v3 ◽

2020 ◽

Author(s):

Xia Li ◽

Jiancong Huang ◽

Xiaohong Chen ◽

Xiaoping Lai ◽

Zizhen Huang ◽

...

Keyword(s):

Western Blot ◽

Chronic Rhinosinusitis ◽

Nasal Polyps ◽

Tissue Remodeling ◽

Tissue Inhibitors Of Metalloproteinases ◽

Tissue Samples ◽

Nasal Tissue ◽

Human Nasal Epithelial Cells ◽

Pathway Activation

Abstract Background: Tissue remodeling is a crucial characteristic of chronic rhinosinusitis (CRS). Imbalance between matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) is crucial for the pathologic tissue remodeling in CRS. Elevation of interleukin (IL)-19 or MMP-9 levels in patients with CRS had been proven in previous studies. Here, we aimed to investigate the role of IL-19 in mediating MMP-9 expression in CRS. Methods: Nasal tissue samples were collected from 45 individuals having chronic rhinosinusitis with nasal polyps (CRSwNP), 24 CRS without nasal polyps (CRSsNP), and 17 controls. Expression of IL-19, its receptors (IL-20R1/IL-20R2), and MMP-9 were investigated using RT-qPCR and Immunofluorescence. Human nasal epithelial cells (HNECs) were stimulated by IL-19; ERK phosphorylation, NF-kB pathway activation, and MMP-9 level were detected by RT-qPCR, ELISA, western blot and Immunofluorescence. We also explored the effect of type1/2/3 cytokines on IL-19 production by RT-qPCR, and western blot. Results: Expression levels of IL-19, its receptors (IL-20R1/IL-20R2), and MMP-9 were increased in nasal tissues from individuals with CRSwNP compared to those with CRSsNP as well as the controls. IL-19 significantly elevated the production of MMP-9 in HNECs. Furthermore, IL-19 could activate the ERK and NF-kB pathways, accompanied by increased MMP-9 production in HNECs. Conversely, both ERK and NF-kB inhibitors significantly attenuated the role of IL-19 in MMP-9 production. siRNA knockdown of IL-20R1 suppressed ERK and NF-kB pathway activation, thereby decreasing MMP-9 expression. IL-13 and IL-17A were found to stimulate IL-19 production in HNECs.Conclusion: IL-19, promoted by IL-13 and IL-17A, contributes to the upregulation of secretion of the tissue remodeling factor MMP-9 in patients with CRS.

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The Role of Relaxin-2 in Tissue Remodeling of Chronic Rhinosinusitis With Nasal Polyps

American Journal of Rhinology and Allergy ◽

10.1177/1945892419843828 ◽

2019 ◽

Vol 33 (5) ◽

pp. 490-499 ◽

Cited By ~ 1

Author(s):

Yimin Li ◽

Guojing Tan ◽

Jie Liu ◽

Xia Ke ◽

Yang Shen ◽

...

Keyword(s):

Western Blot ◽

Chronic Rhinosinusitis ◽

Nasal Polyps ◽

Ex Vivo ◽

Tissue Remodeling ◽

Collagen I ◽

Enzyme Linked Immunosorbent Assay ◽

Type I ◽

Relaxin 2 ◽

Tgf Β1

Background Relaxin is a small peptide hormone that regulates extracellular matrix remodeling and reduces fibrosis in a number of organs. Little is known about its impact on chronic rhinosinusitis with nasal polyps (CRSwNP); thus, we aimed to determine the expression of human H2 relaxin (relaxin-2) and its role in tissue remodeling in CRSwNP. Methods Patients were enrolled and divided into the following groups: CRS with NP (CRSwNP; n = 20), CRS without NP (CRSsNP; n = 20), and controls (n = 15). Tissue samples were analyzed by Masson trichrome staining for collagen, while the location and expression of relaxin-2, transforming growth factor beta 1 (TGF-β1), and phosphorylated (p) Smad2/Smad3 were analyzed by immunohistochemistry and Western blot. The expression of relaxin-2, Smad2, Smad3, and TGF-β1 mRNA was tested by quantitative polymerase chain reaction (qPCR). Ex vivo NP were treated with relaxin-2 (n = 15) or TGF-β1 (n = 15). Collagen type I (collagen I), relaxin-2, and TGF-β1 levels in the culture supernatants were examined by enzyme-linked immunosorbent assay, while pSmad2/Smad3 in culture pellets was analyzed by Western blot, and the expression of Smad2 and Smad3 mRNA was tested by qPCR. Results The collagen, relaxin-2, TGF-β1, and pSmad2/Smad3 protein expression levels were significantly decreased in the CRSwNP group compared with the CRSsNP group ( P < .05). The expression of relaxin-2, Smad2, Smad3, and TGF-β1 mRNA in the CRSsNP group was significantly higher than in the CRSwNP and control groups ( P < .05). Compared with the ex vivo controls, in CRSwNP, the levels of TGF-β1, collagen I, pSmad2/Smad3, Smad2, and Smad3 were markedly decreased after relaxin-2 treatment. However, relaxin-2, collagen I, pSmad2/Smad3, Smad2, and Smad3 were remarkably increased after TGF-β1 treatment. Conclusions The antifibrotic effects of relaxin-2 may play a role in tissue remodeling in CRSwNP, but the detailed mechanism deserves further study.

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IL-19 Induced by IL-13/IL-17A in the Nasal Epithelium of Patients with Chronic Rhinosinusitis Upregulates MMP-9 Expression via ERK/NF-kB Signaling Pathway

10.21203/rs.3.rs-46150/v2 ◽

2020 ◽

Author(s):

Xia Li ◽

Jiancong Huang ◽

Xiaohong Chen ◽

Xiaoping Lai ◽

Zizhen Huang ◽

...

Keyword(s):

Western Blot ◽

Chronic Rhinosinusitis ◽

Nasal Polyps ◽

Tissue Remodeling ◽

Tissue Inhibitors Of Metalloproteinases ◽

Tissue Samples ◽

Nasal Tissue ◽

Human Nasal Epithelial Cells ◽

Pathway Activation

Abstract Background: Tissue remodeling is a crucial characteristic of chronic rhinosinusitis (CRS). Imbalance between matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) is crucial for the pathologic tissue remodeling in CRS. Elevation of interleukin (IL)-19 or MMP-9 levels in patients with CRS had been proven in previous studies. Here, we aimed to investigate the role of IL-19 in mediating MMP-9 expression in CRS. Methods: Nasal tissue samples were collected from 45 individuals having chronic rhinosinusitis with nasal polyps (CRSwNP), 24CRS without nasal polyps (CRSsNP), and 17 controls. Expression of IL-19, its receptors (IL-20R1/IL-20R2), and MMP-9 were investigated using RT-qPCR and Immunofluorescence. Human nasal epithelial cells (HNECs) were stimulated by IL-19; ERK phosphorylation, NF-kB pathway activation, and MMP-9 level were detected by RT-qPCR, ELISA, western blot and Immunofluorescence. We also explored the effect of type1/2/3 cytokines on IL-19 production by RT-qPCR, and western blot. Results: Expression levels of IL-19, its receptors (IL-20R1/IL-20R2), and MMP-9 were increased in nasal tissues from individuals with CRSwNP compared to those with CRSsNP as well as the controls. IL-19 significantly elevated the production of MMP-9 in HNECs. Furthermore, IL-19 could activate the ERK and NF-kB pathways, accompanied by increased MMP-9 production in HNECs. Conversely, both ERK and NF-kB inhibitors significantly attenuated the role of IL-19 in MMP-9 production. siRNA knockdown of IL-20R1 suppressed ERK and NF-kB pathway activation, thereby decreasing MMP-9 expression. IL-13 and IL-17A were found to stimulate IL-19 production in HNECs.Conclusion: IL-19, promoted by IL-13 and IL-17A, contributes to the upregulation of secretion of the tissue remodeling factor MMP-9 in patients with CRS.

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Upregulation of Basonuclin1 Is Associated with p63-Involved Epithelial Barrier Impairment and Type-2 Helper T-cell Inflammation in Chronic Rhinosinusitis with Nasal Polyps

International Archives of Allergy and Immunology ◽

10.1159/000516810 ◽

2021 ◽

pp. 1-12

Author(s):

Yunbo Gao ◽

Jingyun Li ◽

Jian Jiao ◽

Ying Li ◽

Chengshuo Wang ◽

...

Keyword(s):

T Cell ◽

Chronic Rhinosinusitis ◽

Nasal Polyps ◽

Mrna Level ◽

Mucosal Inflammation ◽

Epithelial Barrier ◽

Helper T Cell ◽

Downstream Target ◽

Nasal Tissue

Background: Tumor protein p63 has been shown to be important for epithelial dysfunction, including epithelial barrier defects and mucosal inflammation, in the development of chronic rhinosinusitis with nasal polyps (CRSwNP). Basonuclin1 (BNC1), an epithelial-specific transcriptional factor, is a direct downstream target of p63 and thus might be involved in the pathogenesis of CRSwNP. Objective: We sought to investigate whether BNC1 was associated with p63-mediated epithelial barrier defects and nasal mucosal inflammation in CRSwNP. Methods: Nasal tissue biopsies were obtained from 91 patients to CRSwNP, 49 chronic rhinosinusitis without nasal polyps (CRSsNP) patients, and 28 control subjects. Immunohistochemistry and immunofluorescence staining were used to determine the distribution of BNC1 in tissues and localization in cells, respectively. Quantitative PCR was performed to detect the expression levels of BNC1, TP63, epithelial barrier proteins, and type-2 helper T-cell inflammation-related genes. Results: BNC1 mRNA expression was significantly elevated in the tissues in CRSwNP patients compared with CRSsNP (1.96-fold, p = 0.0003) and control groups (2.40-fold, p < 0.0001). BNC1 staining was strongly positive in the nasal epithelium and co-localized with p63-positive epithelial cells. The expression of BNC1 mRNA was strongly correlated with TP63 mRNA level both in tissue biopsies (r = 0.78, p < 0.0001) and epithelial scrapings (r = 0.97, p < 0.0001). BNC1 expression was also positively correlated with epithelial barrier protein genes (CDH1, CLDN1, CLDN4, TJP1, and TJP2) and epithelial genes involved in TH2 inflammation (IL33, CCL26, CLC, and ALOX15). Conclusions: Overexpression of BNC1 may be associated with increased expression of TP63, and possibly contribute to the epithelial barrier defects and TH2 inflammation in CRSwNP.

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Late Breaking Abstract - Aldo-keto reductases expression correlate corticoids efficacy in vitro and on chronic rhinosinusitis with nasal polyps

10.1183/13993003.congress-2021.pa891 ◽

2021 ◽

Author(s):

Anselm Morell ◽

Javier Milara ◽

Lenka Laštovičková ◽

Inés Roger ◽

Ainhoa García-Lliberós ◽

...

Keyword(s):

Chronic Rhinosinusitis ◽

Nasal Polyps

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