WTAP Promotes the Excessive Proliferation of Airway Smooth Muscle Cells in Asthma by Enhancing AXIN1 Levels Through the Recognition of YTHDF2

Overview

Journal Biochem Genet

Specialty Molecular Biology

Date 2024 Oct 25

PMID 39453546

Authors

Xueli Chen

Li Dai

Affiliations

Soon will be listed here.

Abstract

Asthma is a common chronic respiratory disease in children, the incidence rate of which has increased in recent years. Wilms tumour 1-associated protein (WTAP) is an N6-methyladenosine (m6A) methyltransferase. The purpose of this study was to explore the specific mechanism of WTAP in asthma progression, and clarify the intricate interplay between m6A modifications, WTAP, AXIN1, and their collective impact on airway smooth muscle cells (ASMCs) proliferation in asthma. Platelet-derived growth factor-BB (PDGF-BB)-treated ASMCs were used to establish an asthma model in vitro. The cell phenotype was tested using CCK-8, transwell, and wound healing assays. The expression of the Wnt signalling pathway was detected by western blotting. In addition, the relationship between WTAP/YTDHF2 and AXIN1 was assessed by a double luciferase reporter assay. Actinomycin D treatment and RT‒qPCR assays were performed to determine the mRNA stability of AXIN1. We found that WTAP was significantly increased in PDGF-BB-treated ASMCs. Knockdown of WTAP inhibited the excessive cell viability and migration of ASMCs induced by PDGF-BB. Furthermore, WTAP knockdown increased AXIN1 levels and inhibited the Wnt signalling pathway. Furthermore, WTAP knockdown decreased the m6A levels and enhanced the mRNA stability of AXIN1. WTAP overexpression showed the opposite effect. In addition, YTHDF2 was demonstrated to be the reader that recognizes the WTAP-mediated m6A modification of AXIN1. YTHDF2 knockdown enhanced the mRNA stability of AXIN1 and reversed the effect of WTAP overexpression on PDGF-BB-treated ASMCs. WTAP knockdown inhibited the excessive cell viability and migration of ASMCs by enhancing the m6A levels of AXIN1, which was further recognized by YTHDF2. The upregulation of AXIN1 mediated by the WTAP/YTHDF2 axis further inhibited the Wnt signalling pathway. Our study provides a new method for the treatment of asthma. This work not only deepens our understanding of the molecular underpinnings of asthma but also identifies potential therapeutic targets for the development of novel treatments aimed at inhibiting ASMC proliferation and alleviating asthma symptoms.

References

Camoretti-Mercado B, Lockey R . Airway smooth muscle pathophysiology in asthma. J Allergy Clin Immunol. 2021; 147(6):1983-1995. DOI: 10.1016/j.jaci.2021.03.035. View

Chen Y, Peng C, Chen J, Chen D, Yang B, He B . WTAP facilitates progression of hepatocellular carcinoma via m6A-HuR-dependent epigenetic silencing of ETS1. Mol Cancer. 2019; 18(1):127. PMC: 6704583. DOI: 10.1186/s12943-019-1053-8. View

Dai B, Sun F, Cai X, Li C, Liu H, Shang Y . Significance of RNA N6-Methyladenosine Regulators in the Diagnosis and Subtype Classification of Childhood Asthma Using the Gene Expression Omnibus Database. Front Genet. 2021; 12:634162. PMC: 7982807. DOI: 10.3389/fgene.2021.634162. View

Deng J, Zhang J, Ye Y, Liu K, Zeng L, Huang J . -methyladenosine-Mediated Upregulation of WTAPP1 Promotes WTAP Translation and Wnt Signaling to Facilitate Pancreatic Cancer Progression. Cancer Res. 2021; 81(20):5268-5283. PMC: 9662857. DOI: 10.1158/0008-5472.CAN-21-0494. View

Dixit D, Prager B, Gimple R, Poh H, Wang Y, Wu Q . The RNA m6A Reader YTHDF2 Maintains Oncogene Expression and Is a Targetable Dependency in Glioblastoma Stem Cells. Cancer Discov. 2020; 11(2):480-499. PMC: 8110214. DOI: 10.1158/2159-8290.CD-20-0331. View

Gans M, Gavrilova T . Understanding the immunology of asthma: Pathophysiology, biomarkers, and treatments for asthma endotypes. Paediatr Respir Rev. 2019; 36:118-127. DOI: 10.1016/j.prrv.2019.08.002. View

Abul M, Phipatanakul W . Severe asthma in children: Evaluation and management. Allergol Int. 2019; 68(2):150-157. DOI: 10.1016/j.alit.2018.11.007. View

James A, Noble P, Drew S, Mauad T, Bai T, Abramson M . Airway smooth muscle proliferation and inflammation in asthma. J Appl Physiol (1985). 2018; 125(4):1090-1096. DOI: 10.1152/japplphysiol.00342.2018. View

Jia X, Zhu T, Huang Y, Zeng X, Zhang H, Zhang W . Wnt/β-catenin signaling pathway regulates asthma airway remodeling by influencing the expression of c-Myc and cyclin D1 via the p38 MAPK-dependent pathway. Exp Ther Med. 2019; 18(5):3431-3438. PMC: 6777302. DOI: 10.3892/etm.2019.7991. View

10.

Lan J, Xu B, Shi X, Pan Q, Tao Q . WTAP-mediated N-methyladenosine modification of NLRP3 mRNA in kidney injury of diabetic nephropathy. Cell Mol Biol Lett. 2022; 27(1):51. PMC: 9235192. DOI: 10.1186/s11658-022-00350-8. View

11.

Li V, Ng S, Boersema P, Low T, Karthaus W, Gerlach J . Wnt signaling through inhibition of β-catenin degradation in an intact Axin1 complex. Cell. 2012; 149(6):1245-56. DOI: 10.1016/j.cell.2012.05.002. View

12.

Li Y, Sheng H, Ma F, Wu Q, Huang J, Chen Q . RNA mA reader YTHDF2 facilitates lung adenocarcinoma cell proliferation and metastasis by targeting the AXIN1/Wnt/β-catenin signaling. Cell Death Dis. 2021; 12(5):479. PMC: 8116339. DOI: 10.1038/s41419-021-03763-z. View

13.

Li Z, Zheng Z, Yang P, Lin L, Zhou G, Lv J . WTAP-mediated mA modification of lncRNA DIAPH1-AS1 enhances its stability to facilitate nasopharyngeal carcinoma growth and metastasis. Cell Death Differ. 2022; 29(6):1137-1151. PMC: 9177844. DOI: 10.1038/s41418-021-00905-w. View

14.

He Y, Gallman A, Xie C, Shen Q, Ma J, Wolfreys F . P2RY8 variants in lupus patients uncover a role for the receptor in immunological tolerance. J Exp Med. 2021; 219(1). PMC: 8669517. DOI: 10.1084/jem.20211004. View

15.

Mazzoni S, Fearon E . AXIN1 and AXIN2 variants in gastrointestinal cancers. Cancer Lett. 2014; 355(1):1-8. PMC: 4298141. DOI: 10.1016/j.canlet.2014.09.018. View

16.

Mo B, Li X, Li S, Xiao B, Yang J, Li H . m6A echoes with DNA methylation: Coordinated DNA methylation and gene expression data analysis identified critical m6A genes associated with asthma. Gene. 2022; 828:146457. DOI: 10.1016/j.gene.2022.146457. View

17.

Peng Y, Xu Y, Zhang X, Deng S, Yuan Y, Luo X . A novel protein AXIN1-295aa encoded by circAXIN1 activates the Wnt/β-catenin signaling pathway to promote gastric cancer progression. Mol Cancer. 2021; 20(1):158. PMC: 8642992. DOI: 10.1186/s12943-021-01457-w. View

18.

Qin Y, Li L, Luo E, Hou J, Yan G, Wang D . Role of m6A RNA methylation in cardiovascular disease (Review). Int J Mol Med. 2020; 46(6):1958-1972. PMC: 7595665. DOI: 10.3892/ijmm.2020.4746. View

19.

Tian J, Zhu Y, Rao M, Cai Y, Lu Z, Zou D . N-methyladenosine mRNA methylation of regulates AKT signalling to promote PTEN-deficient pancreatic cancer progression. Gut. 2020; 69(12):2180-2192. DOI: 10.1136/gutjnl-2019-320179. View

20.

Wang J, Lu A . The biological function of m6A reader YTHDF2 and its role in human disease. Cancer Cell Int. 2021; 21(1):109. PMC: 7885220. DOI: 10.1186/s12935-021-01807-0. View