GEOTranscriptomicsHomo sapiens OtherExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE298598GEOGSEfalseMETTL3/YTHDF2 axis inhibits cell proliferation and fibrosis by degrading the N6-methyladenosine modification of TCF7L2-Smad2/Smad3 mRNA in pterygiumBackground: Proliferation and fibrosis of stromal fibroblasts are key mechanisms in pterygium development and recurrence. N6-methyladenosine (m6A) is the most abundant mRNA modification, of which methyltransferase-like 3 (METTL3) is the only catalytic subunit. This study aimed to investigate the regulatory role of METTL3-mediated m6A modification on cell proliferation and fibrosis in pterygium. Methods: Primary human conjunctival fibroblasts (HCFs) and human pterygium fibroblasts (HPFs) were isolated and cultured from fresh conjunctival grafts and pterygium tissues. Adenovirus (ADV) system was used to construct overexpression and silencing of METTL3 in HCFs, combined with TGF-β1 induction, for cell phenotypic analysis. The regulatory role of the METTL3/YTHDF2 axis on target genes (TCF7L2, Smad2, Smad3 and Smad7) was determined using methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA Immunoprecipitation qPCR (RIP-qPCR). The regulatory role of the transcription factors (TCF7L2, Smad2 and Smad3) on the fibrosis genes fibronectin-1 (FN-1) and collagen, type I, alpha 1 (COL1A1) was determined by dual luciferase reporter gene assay. Protein-protein docking (PPD) experiments and co-immunoprecipitation (co-IP) assays were conducted to investigate whether TCF7L2 forms a complex with Smad2/Smad3. Results: HCFs induced by TGF-β1 decreased METTL3 expression, and the reduced METTL3 enhanced HCFs’ proliferation and promoted the expression of fibrotic proteins FN-1 and COL1A1. MeRIP-seq and RIP-qPCR results showed that METTL3-mediated the m6A modification of TCF7L2, Smad2, Smad3 and Smad7 mRNA, was further recognized and degraded by YTHDF2. Dual luciferase reporter gene experiments revealed that TCF7L2, Smad2 and Smad3 is involved in the transcription of genes FN-1 and COL1A1. PPD and co-IP results demonstrate that TCF7L2 is capable of forming a complex with Smad2/Smad3 to regulate the transcription of FN-1 and COL1A1. Conclusion: The METTL3/YTHDF2 axis in pterygium regulates the degradation of the transcriptional complex TCF7L2-Smad2/Smad3 mRNA, promoting the transcription of FN-1 and COL1A1 and the progression of fibrosis. Our study findings untangle the biological characteristics of the METTL3/YTHDF2-regulated transcriptional complex in the pathophysiology of pterygium. Keywords: Pterygium, Fibrosis, m6A, METTL3, YTHDF2, Transcription factors2026/05/29GSE298598GSM9018264GSM9018253GSM9018263GSM9018255GSM9018254GSM9018257GSM9018256GSM9018259GSM9018258GSM9018260GSM9018262GSM901826124676298598Homo sapiens