Home
|
Inbox
|
Search
|
View Abstract
Interferon-Gamma Secreted by Neutrophils Induces Chronic Rhinosinusitis Progression Via Epithelial-To-Mesenchymal (EMT) Transition
Description
.abstract img { width:300px !important; height:auto; display:block; text-align:center; margin-top:10px } .abstract { overflow-x:scroll } .abstract table { width:100%; display:block; border:hidden; border-collapse: collapse; margin-top:10px } .abstract td, th { border-top: 1px solid #ddd; padding: 4px 8px; } .abstract tbody tr:nth-child(even) td { background-color: #efefef; } .abstract a { overflow-wrap: break-word; word-wrap: break-word; }
A1284 - Interferon-Gamma Secreted by Neutrophils Induces Chronic Rhinosinusitis Progression Via Epithelial-To-Mesenchymal (EMT) Transition
Author Block: M. Lee1, D. Kim1, R. Khalmuratova1, Y. Kim2, D. Han1, H. Kim1, D. Kim1, J. Park1, H. Shin1; 1Seoul National University, Seoul, Korea, Republic of, 2Chungnam National University, Daejun, Korea, Republic of.
Rationale: Chronic rhinosinusitis (CRS) is a heterogeneous inflammatory disease of the sinonasal mucosa accompanied by a diverse spectrum of inflammatory cells. Although, numerous etiologic factors associate with nasal polyps (NP), the mechanism underlying NP is not fully understood. Previously, we reported that hypoxia-induced epithelial-to-mesenchymal transition (EMT) is frequently observed in Asian NPs, and HIF-1α could be a therapeutic target for nasal polyposis. However, the nasal epithelial cells in NP patients are not only exposed to hypoxia but also to diverse types or combinations of inflammatory milieu. Furthermore, CRS in Asian patients is characterized mainly by the infiltration of neutrophils and an elevated level of IFN-γ. So, we hypothesized that specific immunologic endotypes could induce or accelerate EMT in nasal epithelial cells, and lead to nasal polyp formation.
Methods: Human nasal epithelial cells (hNEC), RPMI2650 cell lines were used in submerged conditions. We also utilized the ALI culture systems using a normal or polyp derived epithelial cells. Immunoblotting, immunofluorescence and immunohistochemistry were done to evaluate EMT markers and signaling molecules in both submerged and ALI conditions. Additionally, those experiments were conducted in sinonasal tissues from CRS patients with or without NP. Boyden transwell system was utilized to measure the capacity of migration in submerged conditions and TEER amount in ALI conditions. We finally employed the murine NP model to evaluate the importance of IFN-γ-induced signaling pathways.
Results: IFN-γ could most induce EMT, which was confirmed by the spindle-shape of cell morphology, modest cytoskeleton rearrangement, increased migration potential and EMT marker changes among the various type of cytokines. Mechanistically, IFN-γ-induced EMT via ERK and p38 pathway, which were known as non-smad pathway of EMT. Next, we checked IFN-γ and EMT marker levels in human nasal mucosa tissues. IFN-γ expression was upregulated in non-eosinophilic NP mucosa compared with tissues of control, CRS only patients, and eosinophilic NP patients. In addition, this IFN-γ expression was found to correlate with E cadherin (an epithelial marker) loss and α-smooth muscle actin (a mesenchymal marker) expression. Treatment of p38 and ERK inhibitor mixture prevented polyp burden in murine NP model and recapitulate mesenchymal phenotypes of nasal polyp.
Conclusion: IFN-γ induce EMT in hNEC and this process is critically mediated by ERK and p38 pathway. This study shows that IFN-γ-induced EMT is likely to contribute to nasal polyposis in CRS, and suggests that p38 and ERK inhibitors be viewed as a therapeutic target for nasal polyposis.
Author Block: M. Lee1, D. Kim1, R. Khalmuratova1, Y. Kim2, D. Han1, H. Kim1, D. Kim1, J. Park1, H. Shin1; 1Seoul National University, Seoul, Korea, Republic of, 2Chungnam National University, Daejun, Korea, Republic of.
Rationale: Chronic rhinosinusitis (CRS) is a heterogeneous inflammatory disease of the sinonasal mucosa accompanied by a diverse spectrum of inflammatory cells. Although, numerous etiologic factors associate with nasal polyps (NP), the mechanism underlying NP is not fully understood. Previously, we reported that hypoxia-induced epithelial-to-mesenchymal transition (EMT) is frequently observed in Asian NPs, and HIF-1α could be a therapeutic target for nasal polyposis. However, the nasal epithelial cells in NP patients are not only exposed to hypoxia but also to diverse types or combinations of inflammatory milieu. Furthermore, CRS in Asian patients is characterized mainly by the infiltration of neutrophils and an elevated level of IFN-γ. So, we hypothesized that specific immunologic endotypes could induce or accelerate EMT in nasal epithelial cells, and lead to nasal polyp formation.
Methods: Human nasal epithelial cells (hNEC), RPMI2650 cell lines were used in submerged conditions. We also utilized the ALI culture systems using a normal or polyp derived epithelial cells. Immunoblotting, immunofluorescence and immunohistochemistry were done to evaluate EMT markers and signaling molecules in both submerged and ALI conditions. Additionally, those experiments were conducted in sinonasal tissues from CRS patients with or without NP. Boyden transwell system was utilized to measure the capacity of migration in submerged conditions and TEER amount in ALI conditions. We finally employed the murine NP model to evaluate the importance of IFN-γ-induced signaling pathways.
Results: IFN-γ could most induce EMT, which was confirmed by the spindle-shape of cell morphology, modest cytoskeleton rearrangement, increased migration potential and EMT marker changes among the various type of cytokines. Mechanistically, IFN-γ-induced EMT via ERK and p38 pathway, which were known as non-smad pathway of EMT. Next, we checked IFN-γ and EMT marker levels in human nasal mucosa tissues. IFN-γ expression was upregulated in non-eosinophilic NP mucosa compared with tissues of control, CRS only patients, and eosinophilic NP patients. In addition, this IFN-γ expression was found to correlate with E cadherin (an epithelial marker) loss and α-smooth muscle actin (a mesenchymal marker) expression. Treatment of p38 and ERK inhibitor mixture prevented polyp burden in murine NP model and recapitulate mesenchymal phenotypes of nasal polyp.
Conclusion: IFN-γ induce EMT in hNEC and this process is critically mediated by ERK and p38 pathway. This study shows that IFN-γ-induced EMT is likely to contribute to nasal polyposis in CRS, and suggests that p38 and ERK inhibitors be viewed as a therapeutic target for nasal polyposis.
|
Home
|
Inbox
|
Search
|