Epidermal growth factor upregulates expression of MUC5AC via TMEM16A, in chronic rhinosinusitis with nasal polyps

Jian Jiao; Tao Zhang; Yu Zhang; Jingyun Li; Min Wang; Ming Wang; Ying Li; Xiangdong Wang; Luo Zhang

doi:10.1186/s13223-020-00440-2

Epidermal growth factor upregulates expression of MUC5AC via TMEM16A, in chronic rhinosinusitis with nasal polyps

Allergy Asthma Clin Immunol. 2020 May 27:16:40. doi: 10.1186/s13223-020-00440-2. eCollection 2020.

Authors

Jian Jiao^#^{1

2}, Tao Zhang^#^{1

2

3}, Yu Zhang⁴, Jingyun Li^{1

2}, Min Wang^{1

2}, Ming Wang^{1

2}, Ying Li^{1

2}, Xiangdong Wang^{1

2}, Luo Zhang^{1

2}

Affiliations

¹ Department of Otolaryngology, Head and Neck Surgery, Beijing TongRen Hospital, Capital Medical University, Beijing, 100730 People's Republic of China.
² Beijing Key Laboratory of Nasal Diseases, Beijing Institute of Otolaryngology, No. 17, HouGouHuTong, DongCheng District, Beijing, 100005 People's Republic of China.
³ Department of Otolaryngology, Head and Neck Surgery, Tianjin Union Medical Center, Tianjin, 300121 People's Republic of China.
⁴ Department of Otolaryngology, Head and Neck Surgery, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, 264000 People's Republic of China.

^# Contributed equally.

Abstract

Background: Mucus hypersecretion and goblet cell upregulation are common features of chronic rhinosinusitis with nasal polyps (CRSwNP). Although epidermal growth factor (EGF) has been reported to stimulate the expression of MUC5AC, the major macro-molecular constituent of airway mucus, the precise mechanisms underlying the regulation of MUC5AC expression are still not fully understood. The aim of this study therefore was to investigate the role of EGF in regulation of mucin MUC5AC expression and define the involvement of transmembrane protein 16A (TMEM16A) in mediating the EGF-induced mucus overexpression.

Methods: Human nasal epithelial cells (HNECs) derived from tissue of patients with CRSwNP and control subjects were established as air-liquid interface (ALI) cultures. Differentiated cultures were treated with different concentrations of EGF for 4-24 h, and assessed for the expression of TMEM16A and MUC5AC by real-time RT-PCR, Western blotting, ELISA and immunofluorescence. Cultures pretreated for 30 min with T16Ainh-A01 (a specific TMEM16A inhibitor) or LY294002 (a phosphoinositide 3-kinase (PI3K) inhibitor) were also assessed similarly following EGF treatment.

Results: EGF treatment (10-100 ng/ml for 4-24 h) significantly increased the expression of both TMEM16A and MUC5AC mRNA and protein, as well as the percentage of TMEM16A-positive cells, MUC5AC-positive cells and cells coexpressing TMEM16A and MUC5AC in HNECs compared to control non-EGF-treated HNECs. Pretreatment of the HNECs with T16Ainh-A01 and LY294002 attenuated these EGF-induced effects.

Conclusions: This study demonstrated that EGF upregulates the expression of MUC5AC in HNECs from CRSwNP patients. Furthermore, the EGF-mediated regulation of MUC5AC expression is likely to involve a PI3K-TMEM16A signalling pathway in CRSwNP.

Keywords: Chronic rhinosinusitis with nasal polyps; Epidermal growth factor; Human nasal epithelial cells (HNECs); MUC5AC; Phosphoinositide 3-kinase (PI3K); Transmembrane protein 16A (TMEM16A).