METTL3 Regulates the Senescence of Stem Cells From the Apical Papilla by Mediating the m6A Modification of E2F3

INTRODUCTION AND AIMS: We aimed to investigate whether N6-methyladenosine (m⁶A) methylation modification mediated by METTL3 regulates the senescence of SCAPs, and to elucidate the underlying molecular mechanisms. METHODS: Gain- and loss-of-function assays were performed using METTL3 overexpression and knockdown models to evaluate their effects on SCAP senescence and proliferation. To identify downstream targets of METTL3, Me-RIP (methylated RNA immunoprecipitation) high-throughput sequencing was conducted. The functional role of the METTL3-m⁶A-E2F3 axis in SCAP cell cycle progression was further validated via rescue experiments. RESULTS: SCAP senescence was accompanied by reduced m⁶A modification levels and downregulated METTL3 expression. Functional assays demonstrated that METTL3 overexpression attenuated SCAP senescence and enhanced proliferation, whereas METTL3 knockdown exerted the opposite effects. Me-RIP sequencing identified E2F3 as a potential downstream target gene of METTL3 in regulating SCAP senescence and proliferation. Rescue experiments confirmed that the METTL3-m⁶A-E2F3 axis modulated SCAP cell cycle progression, establishing a mechanistic link between METTL3-mediated m⁶A modification and SCAP fate. CONCLUSION: The METTL3-E2F3 axis serves as a critical anti-senescence mechanism in SCAPs under in vitro culture conditions. CLINICAL RELEVANCE: These findings lay the groundwork for the development of novel epigenetic therapies aimed at improving the therapeutic efficacy of SCAPs in regenerative endodontics.