15-Lipoxygenase 1 in nasal polyps promotes CCL26/eotaxin 3 expression through extracellular signal-regulated kinase activation
15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2–high asthma in association with eosinophilia. Chronic rhinosinusitis with nasal polyps (CRSwNP) is also associated with type 2 inflammation and eosinophilia. CCL26/eotaxin 3 has been reported to be regulated by...
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| Published in | Journal of allergy and clinical immunology Vol. 144; no. 5; pp. 1228 - 1241.e9 |
|---|---|
| Main Authors | , , , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
Elsevier Inc
01.11.2019
Elsevier Limited |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0091-6749 1097-6825 1097-6825 |
| DOI | 10.1016/j.jaci.2019.06.037 |
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| Abstract | 15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2–high asthma in association with eosinophilia. Chronic rhinosinusitis with nasal polyps (CRSwNP) is also associated with type 2 inflammation and eosinophilia. CCL26/eotaxin 3 has been reported to be regulated by 15LO1 in lower airway epithelial cells. However, its relation to 15LO1 in patients with CRSwNP or mechanisms for its activation are unclear.
We sought to evaluate 15LO1 and CCL26 expression in nasal epithelial cells (NECs) from patients with CRSwNP and healthy control subjects (HCs) and determine whether 15LO1 regulates CCL26 in NECs through extracellular signal-regulated kinase (ERK) activation.
15LO1, CCL26, and phosphorylated ERK were evaluated in NECs from patients with CRSwNP and HCs. 15LO1/CCL26 and CCL26/cytokeratin 5 were colocalized by means of immunofluorescence. IL-13–stimulated NECs were cultured at an air-liquid interface with or without 15-lipoxygenase 1 gene (ALOX15) Dicer-substrate short interfering RNAs (DsiRNA) transfection, a specific 15LO1 enzymatic inhibitor, and 2 ERK inhibitors. Expression of 15LO1 and CCL26 mRNA and protein was analyzed by using quantitative RT-PCR, Western blotting, and ELISA.
15LO1 expression was increased in nasal polyp (NP) epithelial cells compared with middle turbinate epithelial cells from patients with CRSwNP and HCs. 15LO1 expression correlated with CCL26 expression and colocalized with CCL26 expression in basal cells of the middle turbinate and NPs from patients with CRSwNP. In primary NECs in vitro, IL-13 induced 15LO1 and CCL26 expression. 15LO1 knockdown and inhibition decreased IL-13–induced ERK phosphorylation and CCL26 expression. ERK inhibition (alone) similarly decreased IL-13-induced CCL26. Phosphorylated ERK expression was increased in NECs from CRSwNP subjects and positively correlated with both 15LO1 and CCL26 expression.
15LO1 expression is increased in NP epithelial cells and contributes to CCL26 expression through ERK activation. 15LO1 could be considered a novel therapeutic target for CRSwNP.
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| AbstractList | Background15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2–high asthma in association with eosinophilia. Chronic rhinosinusitis with nasal polyps (CRSwNP) is also associated with type 2 inflammation and eosinophilia. CCL26/eotaxin 3 has been reported to be regulated by 15LO1 in lower airway epithelial cells. However, its relation to 15LO1 in patients with CRSwNP or mechanisms for its activation are unclear.ObjectiveWe sought to evaluate 15LO1 and CCL26 expression in nasal epithelial cells (NECs) from patients with CRSwNP and healthy control subjects (HCs) and determine whether 15LO1 regulates CCL26 in NECs through extracellular signal-regulated kinase (ERK) activation.Methods15LO1, CCL26, and phosphorylated ERK were evaluated in NECs from patients with CRSwNP and HCs. 15LO1/CCL26 and CCL26/cytokeratin 5 were colocalized by means of immunofluorescence. IL-13–stimulated NECs were cultured at an air-liquid interface with or without 15-lipoxygenase 1 gene (ALOX15) Dicer-substrate short interfering RNAs (DsiRNA) transfection, a specific 15LO1 enzymatic inhibitor, and 2 ERK inhibitors. Expression of 15LO1 and CCL26 mRNA and protein was analyzed by using quantitative RT-PCR, Western blotting, and ELISA.Results15LO1 expression was increased in nasal polyp (NP) epithelial cells compared with middle turbinate epithelial cells from patients with CRSwNP and HCs. 15LO1 expression correlated with CCL26 expression and colocalized with CCL26 expression in basal cells of the middle turbinate and NPs from patients with CRSwNP. In primary NECs in vitro, IL-13 induced 15LO1 and CCL26 expression. 15LO1 knockdown and inhibition decreased IL-13–induced ERK phosphorylation and CCL26 expression. ERK inhibition (alone) similarly decreased IL-13-induced CCL26. Phosphorylated ERK expression was increased in NECs from CRSwNP subjects and positively correlated with both 15LO1 and CCL26 expression.Conclusions15LO1 expression is increased in NP epithelial cells and contributes to CCL26 expression through ERK activation. 15LO1 could be considered a novel therapeutic target for CRSwNP. 15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2-high asthma in association with eosinophilia. Chronic rhinosinusitis with nasal polyps (CRSwNP) is also associated with type 2 inflammation and eosinophilia. CCL26/eotaxin 3 has been reported to be regulated by 15LO1 in lower airway epithelial cells. However, its relation to 15LO1 in patients with CRSwNP or mechanisms for its activation are unclear. We sought to evaluate 15LO1 and CCL26 expression in nasal epithelial cells (NECs) from patients with CRSwNP and healthy control subjects (HCs) and determine whether 15LO1 regulates CCL26 in NECs through extracellular signal-regulated kinase (ERK) activation. 15LO1, CCL26, and phosphorylated ERK were evaluated in NECs from patients with CRSwNP and HCs. 15LO1/CCL26 and CCL26/cytokeratin 5 were colocalized by means of immunofluorescence. IL-13-stimulated NECs were cultured at an air-liquid interface with or without 15-lipoxygenase 1 gene (ALOX15) Dicer-substrate short interfering RNAs (DsiRNA) transfection, a specific 15LO1 enzymatic inhibitor, and 2 ERK inhibitors. Expression of 15LO1 and CCL26 mRNA and protein was analyzed by using quantitative RT-PCR, Western blotting, and ELISA. 15LO1 expression was increased in nasal polyp (NP) epithelial cells compared with middle turbinate epithelial cells from patients with CRSwNP and HCs. 15LO1 expression correlated with CCL26 expression and colocalized with CCL26 expression in basal cells of the middle turbinate and NPs from patients with CRSwNP. In primary NECs in vitro, IL-13 induced 15LO1 and CCL26 expression. 15LO1 knockdown and inhibition decreased IL-13-induced ERK phosphorylation and CCL26 expression. ERK inhibition (alone) similarly decreased IL-13-induced CCL26. Phosphorylated ERK expression was increased in NECs from CRSwNP subjects and positively correlated with both 15LO1 and CCL26 expression. 15LO1 expression is increased in NP epithelial cells and contributes to CCL26 expression through ERK activation. 15LO1 could be considered a novel therapeutic target for CRSwNP. 15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2–high asthma in association with eosinophilia. Chronic rhinosinusitis with nasal polyps (CRSwNP) is also associated with type 2 inflammation and eosinophilia. CCL26/eotaxin 3 has been reported to be regulated by 15LO1 in lower airway epithelial cells. However, its relation to 15LO1 in patients with CRSwNP or mechanisms for its activation are unclear. We sought to evaluate 15LO1 and CCL26 expression in nasal epithelial cells (NECs) from patients with CRSwNP and healthy control subjects (HCs) and determine whether 15LO1 regulates CCL26 in NECs through extracellular signal-regulated kinase (ERK) activation. 15LO1, CCL26, and phosphorylated ERK were evaluated in NECs from patients with CRSwNP and HCs. 15LO1/CCL26 and CCL26/cytokeratin 5 were colocalized by means of immunofluorescence. IL-13–stimulated NECs were cultured at an air-liquid interface with or without 15-lipoxygenase 1 gene (ALOX15) Dicer-substrate short interfering RNAs (DsiRNA) transfection, a specific 15LO1 enzymatic inhibitor, and 2 ERK inhibitors. Expression of 15LO1 and CCL26 mRNA and protein was analyzed by using quantitative RT-PCR, Western blotting, and ELISA. 15LO1 expression was increased in nasal polyp (NP) epithelial cells compared with middle turbinate epithelial cells from patients with CRSwNP and HCs. 15LO1 expression correlated with CCL26 expression and colocalized with CCL26 expression in basal cells of the middle turbinate and NPs from patients with CRSwNP. In primary NECs in vitro, IL-13 induced 15LO1 and CCL26 expression. 15LO1 knockdown and inhibition decreased IL-13–induced ERK phosphorylation and CCL26 expression. ERK inhibition (alone) similarly decreased IL-13-induced CCL26. Phosphorylated ERK expression was increased in NECs from CRSwNP subjects and positively correlated with both 15LO1 and CCL26 expression. 15LO1 expression is increased in NP epithelial cells and contributes to CCL26 expression through ERK activation. 15LO1 could be considered a novel therapeutic target for CRSwNP. [Display omitted] 15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2-high asthma in association with eosinophilia. Chronic rhinosinusitis with nasal polyps (CRSwNP) is also associated with type 2 inflammation and eosinophilia. CCL26/eotaxin 3 has been reported to be regulated by 15LO1 in lower airway epithelial cells. However, its relation to 15LO1 in patients with CRSwNP or mechanisms for its activation are unclear.BACKGROUND15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2-high asthma in association with eosinophilia. Chronic rhinosinusitis with nasal polyps (CRSwNP) is also associated with type 2 inflammation and eosinophilia. CCL26/eotaxin 3 has been reported to be regulated by 15LO1 in lower airway epithelial cells. However, its relation to 15LO1 in patients with CRSwNP or mechanisms for its activation are unclear.We sought to evaluate 15LO1 and CCL26 expression in nasal epithelial cells (NECs) from patients with CRSwNP and healthy control subjects (HCs) and determine whether 15LO1 regulates CCL26 in NECs through extracellular signal-regulated kinase (ERK) activation.OBJECTIVEWe sought to evaluate 15LO1 and CCL26 expression in nasal epithelial cells (NECs) from patients with CRSwNP and healthy control subjects (HCs) and determine whether 15LO1 regulates CCL26 in NECs through extracellular signal-regulated kinase (ERK) activation.15LO1, CCL26, and phosphorylated ERK were evaluated in NECs from patients with CRSwNP and HCs. 15LO1/CCL26 and CCL26/cytokeratin 5 were colocalized by means of immunofluorescence. IL-13-stimulated NECs were cultured at an air-liquid interface with or without 15-lipoxygenase 1 gene (ALOX15) Dicer-substrate short interfering RNAs (DsiRNA) transfection, a specific 15LO1 enzymatic inhibitor, and 2 ERK inhibitors. Expression of 15LO1 and CCL26 mRNA and protein was analyzed by using quantitative RT-PCR, Western blotting, and ELISA.METHODS15LO1, CCL26, and phosphorylated ERK were evaluated in NECs from patients with CRSwNP and HCs. 15LO1/CCL26 and CCL26/cytokeratin 5 were colocalized by means of immunofluorescence. IL-13-stimulated NECs were cultured at an air-liquid interface with or without 15-lipoxygenase 1 gene (ALOX15) Dicer-substrate short interfering RNAs (DsiRNA) transfection, a specific 15LO1 enzymatic inhibitor, and 2 ERK inhibitors. Expression of 15LO1 and CCL26 mRNA and protein was analyzed by using quantitative RT-PCR, Western blotting, and ELISA.15LO1 expression was increased in nasal polyp (NP) epithelial cells compared with middle turbinate epithelial cells from patients with CRSwNP and HCs. 15LO1 expression correlated with CCL26 expression and colocalized with CCL26 expression in basal cells of the middle turbinate and NPs from patients with CRSwNP. In primary NECs in vitro, IL-13 induced 15LO1 and CCL26 expression. 15LO1 knockdown and inhibition decreased IL-13-induced ERK phosphorylation and CCL26 expression. ERK inhibition (alone) similarly decreased IL-13-induced CCL26. Phosphorylated ERK expression was increased in NECs from CRSwNP subjects and positively correlated with both 15LO1 and CCL26 expression.RESULTS15LO1 expression was increased in nasal polyp (NP) epithelial cells compared with middle turbinate epithelial cells from patients with CRSwNP and HCs. 15LO1 expression correlated with CCL26 expression and colocalized with CCL26 expression in basal cells of the middle turbinate and NPs from patients with CRSwNP. In primary NECs in vitro, IL-13 induced 15LO1 and CCL26 expression. 15LO1 knockdown and inhibition decreased IL-13-induced ERK phosphorylation and CCL26 expression. ERK inhibition (alone) similarly decreased IL-13-induced CCL26. Phosphorylated ERK expression was increased in NECs from CRSwNP subjects and positively correlated with both 15LO1 and CCL26 expression.15LO1 expression is increased in NP epithelial cells and contributes to CCL26 expression through ERK activation. 15LO1 could be considered a novel therapeutic target for CRSwNP.CONCLUSIONS15LO1 expression is increased in NP epithelial cells and contributes to CCL26 expression through ERK activation. 15LO1 could be considered a novel therapeutic target for CRSwNP. |
| Author | Wenzel, Sally E. Moore, John A. Li, Zhipeng Zhao, Jinming Zhou, Xiuxia Zhang, Weitian Yin, Shankai Goldschmidt, Ezequiel Lee, Stella E. Trudeau, John B. Di, Y. Peter Deng, Yanhan Zeng, Ming Chu, Hongwei Liu, Zheng |
| Author_xml | – sequence: 1 givenname: Zhipeng surname: Li fullname: Li, Zhipeng organization: Department of Otolaryngology–Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai – sequence: 2 givenname: Ming surname: Zeng fullname: Zeng, Ming organization: Department of Otolaryngology-Head and Neck Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China – sequence: 3 givenname: Yanhan surname: Deng fullname: Deng, Yanhan organization: Department of Respiratory and Critical Care Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China – sequence: 4 givenname: Jinming surname: Zhao fullname: Zhao, Jinming organization: University of Pittsburgh Asthma Institute@UPMC, Department of Environmental and Occupational Health, Graduate School of Public Health, Pittsburgh, Pa – sequence: 5 givenname: Xiuxia surname: Zhou fullname: Zhou, Xiuxia organization: University of Pittsburgh Asthma Institute@UPMC, Department of Environmental and Occupational Health, Graduate School of Public Health, Pittsburgh, Pa – sequence: 6 givenname: John B. surname: Trudeau fullname: Trudeau, John B. organization: University of Pittsburgh Asthma Institute@UPMC, Department of Environmental and Occupational Health, Graduate School of Public Health, Pittsburgh, Pa – sequence: 7 givenname: Ezequiel surname: Goldschmidt fullname: Goldschmidt, Ezequiel organization: Department of Neurological Surgery, University of Pittsburgh Medical Center, Pittsburgh, Pa – sequence: 8 givenname: John A. surname: Moore fullname: Moore, John A. organization: Department of Otolaryngology–Head and Neck Surgery, University of Pittsburgh Medical Center, Mercy Hospital, Pittsburgh, Pa – sequence: 9 givenname: Hongwei surname: Chu fullname: Chu, Hongwei organization: Department of Medicine, National Jewish Health, Denver, Colo – sequence: 10 givenname: Weitian surname: Zhang fullname: Zhang, Weitian organization: Department of Otolaryngology–Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai – sequence: 11 givenname: Shankai surname: Yin fullname: Yin, Shankai organization: Department of Otolaryngology–Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai – sequence: 12 givenname: Zheng surname: Liu fullname: Liu, Zheng organization: Department of Otolaryngology-Head and Neck Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China – sequence: 13 givenname: Y. Peter surname: Di fullname: Di, Y. Peter organization: University of Pittsburgh Asthma Institute@UPMC, Department of Environmental and Occupational Health, Graduate School of Public Health, Pittsburgh, Pa – sequence: 14 givenname: Stella E. surname: Lee fullname: Lee, Stella E. email: lees6@upmc.edu organization: Department of Otolaryngology–Head and Neck Surgery, University of Pittsburgh Medical Center, Mercy Hospital, Pittsburgh, Pa – sequence: 15 givenname: Sally E. surname: Wenzel fullname: Wenzel, Sally E. email: swenzel@pitt.edu organization: University of Pittsburgh Asthma Institute@UPMC, Department of Environmental and Occupational Health, Graduate School of Public Health, Pittsburgh, Pa |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/31301373$$D View this record in MEDLINE/PubMed |
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| Keywords | extracellular signal-regulated kinase NP DMSO IHC ALOX15 HRP AERD PEBP1 CRSwNP WB epithelial cell CK5 Ct SNP UPMC nasal polyps KD GUSB 15-HpETE T2 AA 15LO1 DsiRNA MT qRT-PCR 15-HETE IT NEC 15-Lipoxygenase 1 IL-4R PE HAEC chronic rhinosinusitis HC GAPDH pERK ALI CCL26 ERK tERK |
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| PublicationDate | November 2019 2019-11-00 20191101 |
| PublicationDateYYYYMMDD | 2019-11-01 |
| PublicationDate_xml | – month: 11 year: 2019 text: November 2019 |
| PublicationDecade | 2010 |
| PublicationPlace | United States |
| PublicationPlace_xml | – name: United States – name: St. Louis |
| PublicationTitle | Journal of allergy and clinical immunology |
| PublicationTitleAlternate | J Allergy Clin Immunol |
| PublicationYear | 2019 |
| Publisher | Elsevier Inc Elsevier Limited |
| Publisher_xml | – name: Elsevier Inc – name: Elsevier Limited |
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| Snippet | 15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2–high asthma in association with eosinophilia. Chronic rhinosinusitis... 15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2-high asthma in association with eosinophilia. Chronic rhinosinusitis... Background15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2–high asthma in association with eosinophilia. Chronic... |
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| SubjectTerms | 15-Lipoxygenase 1 Adult Antigens Arachidonate 15-Lipoxygenase - genetics Arachidonate 15-Lipoxygenase - metabolism Asthma Basal cells Bone CCL26 Cells, Cultured Chemokine CCL26 - metabolism Chronic Disease chronic rhinosinusitis Cytokeratin Enzyme Activation Enzyme-linked immunosorbent assay Eosinophilia Eotaxin epithelial cell Epithelial cells Extracellular signal-regulated kinase Extracellular Signal-Regulated MAP Kinases - metabolism Female Hospitals Humans Immunofluorescence Immunoglobulins Interleukin 13 Kinases Lipoxygenase Male Methods Microscopy Middle Aged mRNA nasal polyps Nasal Polyps - complications Nasal Polyps - metabolism Nose Otolaryngology Phosphorylation Polymerase chain reaction Polyps Proteins Respiratory Mucosa - metabolism Respiratory Mucosa - pathology Respiratory tract Rhinitis Rhinitis - complications Rhinitis - metabolism Rhinosinusitis RNA, Small Interfering - genetics Sinusitis Sinusitis - complications Sinusitis - metabolism Therapeutic applications Transfection Turbinates - metabolism Up-Regulation Western blotting |
| Title | 15-Lipoxygenase 1 in nasal polyps promotes CCL26/eotaxin 3 expression through extracellular signal-regulated kinase activation |
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