KDM6B-Mediated HADHA Demethylation/Lactylation Regulates Cementogenesis

Abstract

Cementum, a bone-like tissue, is an essential component of periodontium, and periodontitis can lead to degenerative changes in the cementum, eventually resulting in tooth loss. The therapeutic strategy for advanced periodontitis is to achieve periodontal regeneration, of which cementum regeneration is a key criterion. Cementoblasts are responsible for cementogenesis, and their mineralization counts in cementum regeneration. However, research is still limited. Thus, novel treatment targets are required. The expression levels of lysine (K)–specific demethylase 6B (KDM6B), fatty acid oxidation (FAO), and cementogenic markers were detected by quantitative polymerase chain reaction, Western blot, immunofluorescence, and immunohistochemical assays. FAO levels were analyzed by assay kit. In vivo, injection of GSK-J4 into mice detected the influence of KDM6B on cementum formation. Chromatin immunoprecipitation sequencing, transcriptomic RNA sequencing, subsequent chromatin immunoprecipitation–quantitative polymerase chain reaction and overexpression of HADHA (hydroxyacyl-coA dehydrogenase trifunctional multienzyme complex subunit alpha) elucidated the KDM6B-Hadha axis. Global lactylation was detected by Western blot. Lactylation proteomics clarified the modified sites of HADHA. Mutating these sites and applying coimmunoprecipitation confirmed their significance. Knockdown of Kdm6b was utilized to assess its regulation on the lactylation of HADHA, FAO, and mineralization levels. FAO and KDM6B expression was elevated during cementoblast mineralization. KDM6B targeted Hadha and activated its transcription, thereby increasing FAO levels and promoting mineralization. Lactylation occurred in the process of mineralization, and KDM6B could regulate the lactylation of HADHA to promote FAO and mineralization. Overexpression of Hadha and the addition of lactate sodium could rescue the inhibition of mineralization by knockdown of Kdm6b. In summary, during cementoblast mineralization, KDM6B regulates HADHA by mediating histone demethylation and lactylation, thereby upregulating FAO and thus promoting mineralization.