基于KDM4B-DLX3通路的颌骨调控牙齿早期发育的表观遗传学机制研究

牙齿发育早期的信号调控机制是功能性牙齿发育及再生的关键。普遍观点认为，牙齿发育的全部信息都编写于牙胚的上皮和间充质基因中，几乎不再接受外界的信号调控。我们前期在模型动物“小型猪”中却证实，下颌骨通过在特定时期释放携带ssc-mir-133b信号的外泌体，调控牙胚的形态发生；进一步研究还发现，基于组蛋白去甲基化酶KDM4B/转录因子Dlx3的表观遗传学机制可能也参与了牙胚发育早期颌骨与牙胚间的通讯。据此，本课题拟采用RNA-Seq、ChIP、Co-IP、ChIP-Seq等方法，筛选出颌骨分泌的、可作用于KDM4B的非编码miRNA，验证是否存在KDM4B对牙发育关键转录因子Dlx3的调控作用，探索在颌骨-牙胚通讯中的“miRNA-KDM4B-转录因子轴”。本研究的预期结果有望对经典的、牙齿发育完全“自洽”的理论做出一定的修正与完善，为基于发育模式的牙再生提供新的理论依据。

The signaling regulation mechanism in the early stage of tooth development is crucial to functional tooth development and regeneration. Classical theory holds that all information is written in the epithelial and mesenchymal genes of the tooth germ, and is almost no longer affected by the external microenvironment. However, we have previously demonstrated that mandible regulated the morphogenesis of the tooth germ by releasing exosomes with ssc-mir-133b at specific times in miniature pigs; further studies have also found that histone demethylation might be involved. The epigenetic mechanism of KDM4B/Dlx3 seems to play an important role in the communication between mandible and tooth germ in the early stage of tooth development. Therefore, in the current study, RNA-Seq, ChIP, Co-IP, ChIP-Seq are used to search the non-coding miRNAs secreted by mandible and verify KDM4B/Dlx3 in the regulation of tooth development. The expected results of this study are expected to revise and improve the classical theory of self-consistency in tooth development, and provide a new theoretical basis for tooth regeneration based on developmental model.