Increased CYR61 expression activates CCND1/c-Myc pathway to promote nasal epithelial cells proliferation in chronic rhinosinusitis with nasal polyps

Chunyu Luo; Ying Zhu; Jiayao Zhou; Xiwen Sun; Shiyao Zhang; Shaolin Tan; Zhipeng Li; Hai Lin; Weitian Zhang

doi:10.1016/j.clim.2023.109235

Increased CYR61 expression activates CCND1/c-Myc pathway to promote nasal epithelial cells proliferation in chronic rhinosinusitis with nasal polyps

Clin Immunol. 2023 Feb:247:109235. doi: 10.1016/j.clim.2023.109235. Epub 2023 Jan 18.

Authors

Chunyu Luo¹, Ying Zhu¹, Jiayao Zhou¹, Xiwen Sun¹, Shiyao Zhang¹, Shaolin Tan², Zhipeng Li¹, Hai Lin³, Weitian Zhang⁴

Affiliations

¹ Department of Otolaryngology-Head and Neck Surgery, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China; Otolaryngological Institute, Shanghai Jiao Tong University, Shanghai, China; Shanghai Key Laboratory of Sleep Disordered Breathing, Shanghai, China.
² Department of Otolaryngology-Head and Neck Surgery, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China; Otolaryngological Institute, Shanghai Jiao Tong University, Shanghai, China; Shanghai Key Laboratory of Sleep Disordered Breathing, Shanghai, China; Postgraduate Training Base of Shanghai Sixth People's Hospital, Jinzhou Medical University, Shanghai, China.
³ Department of Otolaryngology-Head and Neck Surgery, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China; Otolaryngological Institute, Shanghai Jiao Tong University, Shanghai, China; Shanghai Key Laboratory of Sleep Disordered Breathing, Shanghai, China. Electronic address: hailin@sjtu.edu.cn.
⁴ Department of Otolaryngology-Head and Neck Surgery, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China; Otolaryngological Institute, Shanghai Jiao Tong University, Shanghai, China; Shanghai Key Laboratory of Sleep Disordered Breathing, Shanghai, China. Electronic address: drzhangwt@163.com.

PMID: 36681101
DOI: 10.1016/j.clim.2023.109235

Abstract

Purpose: Chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP) is a chronic sinonasal inflammatory disease characterized histologically by hyperplastic nasal epithelium and epithelial cells proliferation. Cysteine-rich angiogenic inducer 61 (CYR61) acts as a positive regulator of cell cycle process. Cyclin D1 (CCND1) and c-Myc play key roles in the processes of cell cycle and cell growth. The purpose of our research was to explore the expression and roles of CYR61, CCND1 and c-Myc in CRSwNP.

Methods: FeaturePlot and vlnPlot functions embedded in the seurat package (version 4.1.1) of R software (version 4.2.0) were applied to explore the cellular distribution of CYR61, CCND1 and c-Myc in the single-cell RNA sequencing (scRNA-seq) dataset of nasal tissue samples. CYR61, CCND1 and c-Myc immunolabeling and mRNA levels in nasal tissue samples were assessed by immunohistochemistry and real-time PCR. Co-localization of CYR61, CCND1 and c-Myc with basal epithelial cell marker P63 was assayed using double-label immunofluorescence staining. Furthermore, we collected and cultured human nasal epithelial cells (HNEC) to assess the regulation and role of CYR61 in vitro study.

Results: CYR61, CCND1 and c-Myc were primarily expressed by nasal epithelial cells. Significant upregulation of CYR61, CCND1 and c-Myc positive cells and increased levels of CYR61, CCND1 and c-Myc mRNA were found in nasal polyps in comparison to control samples. Of note, CYR61 mRNA and protein levels were altered by SEB, LPS, IFN-γ, IL-13, IL-17A and TGF-β1 in HNEC. In addition, CYR61 intervention could increase CCND1 and c-Myc mRNA and protein levels to promote HNEC proliferation, and siRNA against ITGA2 (si-ITGA2) could reverse CYR61 induced upregulation of CCND1 and c-Myc mRNA and protein levels in HNEC and cell proliferation of HNEC.

Conclusions: CYR61, CCND1 and c-Myc were primarily expressed by epithelial cells in nasal mucosa. CYR61, CCND1 and c-Myc expression levels were increased in CRSwNP compared with controls. CYR61 could interact with ITGA2 to enhance HNEC proliferation via upregulating CCND1 and c-Myc levels in the HNEC, leading to hyperplastic nasal epithelium in CRSwNP.

Keywords: C-Myc; CCND1; CYR61; Chronic rhinosinusitis; Human nasal epithelial cells; Nasal polyp.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Proliferation
Chronic Disease
Cyclin D1 / genetics
Cyclin D1 / metabolism
Cysteine-Rich Protein 61* / metabolism
Epithelial Cells / metabolism
Humans
Nasal Mucosa / metabolism
Nasal Polyps* / metabolism
RNA, Messenger / metabolism
Rhinitis* / metabolism

Substances

CCND1 protein, human
Cyclin D1
RNA, Messenger
CCN1 protein, human
Cysteine-Rich Protein 61