METTL3-mediated pre-miR-665/DLX3 m6A methylation facilitates the committed differentiation of stem cells from apical papilla

Abstract

Methyltransferase-like 3 (METTL3) is a crucial element of N6-methyladenosine (m6A) modifications and has been extensively studied for its involvement in diverse biological and pathological processes. In this study, we explored how METTL3 affects the differentiation of stem cells from the apical papilla (SCAPs) into odonto/osteoblastic lineages through gain- and loss-of-function experiments. The m6A modification levels were assessed using m6A dot blot and activity quantification experiments. In addition, we employed Me-RIP microarray experiments to identify specific targets modified by METTL3. Furthermore, we elucidated the molecular mechanism underlying METTL3 function through dual-luciferase reporter gene experiments and rescue experiments. Our findings indicated that METTL3+/− mice exhibited significant root dysplasia and increased bone loss. The m6A level and odonto/osteoblastic differentiation capacity were affected by the overexpression or inhibition of METTL3. This effect was attributed to the acceleration of pre-miR-665 degradation by METTL3-mediated m6A methylation in cooperation with the “reader” protein YTHDF2. Additionally, the targeting of distal-less homeobox 3 (DLX3) by miR-665 and the potential direct regulation of DLX3 expression by METTL3, mediated by the “reader” protein YTHDF1, were demonstrated. Overall, the METTL3/pre-miR-665/DLX3 pathway might provide a new target for SCAP-based tooth root/maxillofacial bone tissue regeneration. Dentition defects, or missing teeth, can cause long-term health issues and are typically treated with implant denture restoration, a method that often has complications. Recent studies suggest using dental stem cells for treatment. A study by Nanjing Medical University investigated the role of a molecule, METTL3, in the development of stem cells from the apical papilla (SCAPs), a type of dental stem cell. They discovered that METTL3 helps SCAPs develop into odonto/osteogenic cells, important for tooth and bone growth. This implies that METTL3 could enhance treatments for missing teeth. The study also found a regulatory network involving METTL3 and other molecules, which could be significant in future research. This summary was initially drafted using artificial intelligence, then revised and fact-checked by the author.